BACKGROUND Knee osteoarthritis(OA)is a degenerative joint disease traditionally viewed through the lens of cartilage degradation.However,emerging evidence positions subchondral bone pathology-particularly bone marrow ...BACKGROUND Knee osteoarthritis(OA)is a degenerative joint disease traditionally viewed through the lens of cartilage degradation.However,emerging evidence positions subchondral bone pathology-particularly bone marrow lesions(BMLs)-as a key contributor to pain,progression,and structural deterioration.Mesenchymal stem cell exhaustion within the osteoarthritic subchondral zone further impairs intrinsic repair mechanisms,reinforcing the rationale for biologic interventions.AIM To evaluate the clinical efficacy of bone marrow aspirate concentrate(BMAC)therapy for knee OA,comparing subchondral vs intra-articular delivery routes,and elucidating the therapeutic impact on symptom relief and structural preservation.METHODS Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines,five clinical studies were included-comprising three randomized controlled trials and two prospective cohorts-with pooled data from 298 knees.Data on functional outcomes,imaging findings,and progression to total knee arthroplasty(TKA)were extracted and qualitatively synthesized.RESULTS Subchondral BMAC injections demonstrated superior improvements compared to intra-articular injection or placebo:Knee Injury and Osteoarthritis Outcome Score improved from 49.1±1.9 to 61.2±6.3 at 12 months(P<0.05),Knee Society Score increased from 57±12 to 87.3±12 at two years,and Western Ontario and McMaster Universities Arthritis Index scores showed significant improvement favoring combined approaches.Magnetic resonance imaging analyses revealed mean BML volume regression of 2.1 cm3,with 80%of knees avoiding TKA over 13-year follow-up.Magnetic resonance imaging analyses revealed regression of BMLs and increased cartilage preservation in subchondral-treated knees.Long-term data indicated delayed progression to TKA and biomechanical improvements(e.g.,Hip-Knee-Ankle angle correction).No major adverse events were reported.CONCLUSION Targeting subchondral bone with BMAC addresses underlying OA pathology and may offer disease-modifying potential beyond symptom relief.These findings support a paradigm shift toward whole-joint biologic therapy,positioning the subchondral matrix as a therapeutic epicenter in OA management.展开更多
AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated...AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated cirrhosis were included. Approximately 200 mL of the bone marrow of the patients was aspirated, and CD34+ stem cells were selected. Between 3 to 10 million CD34+ cells were isolated. The cells were slowly infused through the hepatic artery of the patients.RESULTS: Patient 1 showed marginal improvement in serum albumin and no significant changes in other test results. In patient 2 prothrombin time was decreased; however, her total bilirubin, serum creatinine, and Model of End-Stage Liver Disease (MELD) score worsened at the end of follow up. In patient 3 there was improvement in serum albumin, porthrombin time (PT), and MELD score. Patient 4 developed radiocontrast nephropathy after the procedure, and progressed to type 1 hepatorenal syndrome and died of liver failure a few days later. Because of the major side effects seen in the last patient, the trial was prematurely stopped.CONCLUSION: Infusion of CD34+ stem cells through the hepatic artery is not safe in decompensated cirrhosis. Radiocontrast nephropathy and hepatorenal syndrome could be major side effects. However, this study doesnot preclude infusion of CD34+ stem cells through other routes.展开更多
Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase dige...Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.展开更多
Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that cita...Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that citalopram, a selective serotonin reuptake inhibitor, can promote the neuronal differentiation of adult bone marrow mesenchymal stem cells. Citalopram strongly enhanced neuronal characteristics of the cells derived from bone marrow mesenchymal stem cells. The rate of cell death was decreased in citalopram-treated bone marrow mesenchymal stem cells than in control cells in neurobasal medium. In addition, the cumulative population doubling level of the citalopram-treated cells was signiifcantly increased compared to that of control cells. Also BrdU incorporation was elevated in citalopram-treated cells. These ifndings suggest that citalopram can improve the neuronal-like cell differentiation of bone marrow mesenchymal stem cells by increasing cell proliferation and survival while maintaining their neuronal characteristics.展开更多
Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. ...Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. Relatively fewcomparative studies have been performed to assess the effects of cryopreservationmethods on these stem cells. Dimethyl sulfoxide (DMSO) has been a key agent forthe development of cryobiology and has been used universally for cryopreservation.However, the use of DMSO has been associated with in vitro and in vivotoxicity and has been shown to affect many cellular processes due to changes inDNA methylation and dysregulation of gene expression. Despite studies showingthat DMSO may affect cell characteristics, DMSO remains the CPA of choice, bothin a research setting and in the clinics. However, numerous alternatives to DMSOhave been shown to hold promise for use as a CPA and include albumin,trehalose, sucrose, ethylene glycol, polyethylene glycol and many more. Here, wewill discuss the use, advantages and disadvantages of these CPAs for cryopreservationof different types of stem cells, including hematopoietic stem cells,mesenchymal stromal/stem cells and induced pluripotent stem cells.展开更多
Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell t...Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell type and its physiological state.Accordingly,EVs derived from healthy cells may have a comparable therapeutic potential as cells themselves.展开更多
BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(P...BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(PD-L1),which determines the clinical therapeutic efficacy of MSCs for inflammatory or immune diseases.In MSCs,interferon-gamma(IFN-γ)is a key inducer of PD-L1 expression,which is synergistically enhanced by tumor necrosis factor-alpha(TNF-α);however,the underlying mechanism is unclear.AIM To reveal the mechanism of pretreated MSCs express high PD-L1 and explore the application of pretreated MSCs in ulcerative colitis.METHODS We assessed PD-L1 expression in human umbilical-cord-derived MSCs(hUC-MSCs)induced by IFN-γand TNF-α,alone or in combination.Additionally,we performed signal pathway inhibitor experiments as well as RNA interference experiments to elucidate the molecular mechanism by which IFN-γalone or in combination with TNF-αinduces PD-L1 expression.Moreover,we used luciferase reporter gene experiments to verify the binding sites of the transcription factors of each signal transduction pathway to the targeted gene promoters.Finally,we evaluated the immunosuppressive capacity of hUC-MSCs treated with IFN-γand TNF-αin both an in vitro mixed lymphocyte culture assay,and in vivo in mice with dextran sulfate sodium-induced acute colitis.RESULTS Our results suggest that IFN-γinduction alone upregulates PD-L1 expression in hUC-MSCs while TNF-αalone does not,and that the co-induction of IFN-γand TNF-αpromotes higher expression of PD-L1.IFN-γinduces hUCMSCs to express PD-L1,in which IFN-γactivates the JAK/STAT1 signaling pathway,up-regulates the expression of the interferon regulatory factor 1(IRF1)transcription factor,promotes the binding of IRF1 and the PD-L1 gene promoter,and finally promotes PD-L1 mRNA.Although TNF-αalone did not induce PD-L1 expression in hUCMSCs,the addition of TNF-αsignificantly enhanced IFN-γ-induced JAK/STAT1/IRF1 activation.TNF-αupregulated IFN-γreceptor expression through activation of the nuclear factor kappa-B signaling pathway,which significantly enhanced IFN-γsignaling.Finally,co-induced hUC-MSCs have a stronger inhibitory effect on lymphocyte proliferation,and significantly ameliorate weight loss,mucosal damage,inflammatory cell infiltration,and up-regulation of inflammatory factors in colitis mice.CONCLUSION Overall,our results suggest that IFN-γand TNF-αenhance both the immunosuppressive ability of hUC-MSCs and their efficacy in ulcerative colitis by synergistically inducing high expression of PD-L1.展开更多
Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in...Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in the hippocampus and ventricular subventricular zone lining the lateral ventricle(Olpe and Jessberger,2022).Adult NSCs in rodents are preserved throughout life and continuously produce new neurons that integrate into the pre-existing neuronal network.展开更多
CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentia...CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentiation from MSCs, especially from human MSCs (hUC-MSCs) are still unclear. In the current study, the effect of CD34 antibody on the CD34+ cell differentiation from human umbilical cord (UC)-derived MSCs (hUC-MSCs) is determined. The results have demonstrated that the expression of cd34 protein is significantly increased in hUC-MSCs treated with CD34 antibody. In addition, the cell proliferation is increased in hUC-MSCs after treatment with CD34 antibody. Moreover, the expression of PI3K, AKT, p-AKT proteins, which are signaling molecules related to stem cell differentiation, is increased by CD34 antibody. The results suggest that CD34 antibody could promote the differentiation of hUC-MSCs into CD34+ cells and PI3K/AKT may be involved in this important process.展开更多
Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into th...Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.展开更多
Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat th...Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat through optimization of the cell source and organization process.Results We engineered fibroblasts into myogenic cells via non-viral introduction of the MYOD1 gene,avoiding viral methods for safety.After confirming the stable derivation of myogenic cells,we combined knockout(KO)of MSTN,a negative regulator of myogenesis,with MYOD1-mediated myogenesis to improve cultivated meat production.Primary cells from MSTN KO cattle exhibited enhanced myogenic potential.Additionally,when tested in immortalized fibroblasts,myostatin treatment reduced MYOD1-induced myogenesis in two-dimensional cultures,while MSTN knockout increased it.To achieve muscle-like cell alignment,we employed digital light processing(DLP)-based three-dimensional(3D)bioprinting to organize cells into 3D groove-shaped hydrogels.These bioactive hydrogels supported stable cell proliferation and significantly improved muscle cell alignment.Upon differentiation into myotubes,the cells demonstrated an ordered alignment,particularly the MSTN KO cells,which showed highly efficient differentiation.Conclusions The integration of genetic modification and advanced DLP 3D bioprinting with groove-patterned hydrogels provides an effective strategy for producing high-quality,muscle-aligned cultivated meat.展开更多
“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging...“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging of the human brain is characterized by a progressive decline of its functional capacity;this decline however varies widely,and cognitive longevity differs substantially between individuals.展开更多
Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of pr...Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix(ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. Methods: Liver tissues from both control(normal) and carbon tetrachloride(CCl_(4))-induced fibrotic rats were decellularized using sodium dodecyl sulfate(SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Microparticles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues(MTs) comprising Huh-7 cells, human umbilical vein endothelial cells(HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. Results: Fibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition(EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. Conclusions: Our findings suggest a regulatory role for the fibrotic matrix in promoting cancerous phenotype, which could potentially accelerate the progression of malignancy in the liver.展开更多
Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimul...Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimulation techniques, fail to induce a repair tissue with the same functional and mechanical properties of native hyaline cartilage. Osteochondral transplantation is considered an effective treatment option but is as-sociated with some disadvantages, including donor-site morbidity, tissue supply limitation, unsuitable mechani-cal properties and thickness of the obtained tissue. Although autologous chondrocyte implantation results in reasonable repair, it requires a two-step surgical pro-cedure. Moreover, chondrocytes expanded in culture gradually undergo dedifferentiation, so lose morpho-logical features and specialized functions. In the search for alternative cells, scientists have found mesenchymal stem cells(MSCs) to be an appropriate cellular mate-rial for articular cartilage repair. These cells were origi-nally isolated from bone marrow samples and further investigations have revealed the presence of the cells in many other tissues. Furthermore, chondrogenic dif-ferentiation is an inherent property of MSCs noticedat the time of the cell discovery. MSCs are known to exhibit homing potential to the damaged site at which they differentiate into the tissue cells or secrete a wide spectrum of bioactive factors with regenerative proper-ties. Moreover, these cells possess a considerable im-munomodulatory potential that make them the general donor for therapeutic applications. All of these topics will be discussed in this review.展开更多
Acupuncture has been commonly used as an adjuvant therapy or monotherapy in the treatment of Parkinson's disease in China and in other countries.Animal studies have consistently show that this treatment is both ne...Acupuncture has been commonly used as an adjuvant therapy or monotherapy in the treatment of Parkinson's disease in China and in other countries.Animal studies have consistently show that this treatment is both neuroprotective, protecting dopaminergic neurons from degeneration and also restorative, restoring tyrosine hydroxylase positive dopaminergic terminals in striatum, resulting in improvements in motor performance in animal models of Parkinsonism. Studies show that this protection is mediated through the same common mechanisms as other neuroprotective agents, including anti-oxidative stress, anti-inflammatory and anti-apoptotic pathways at molecular and cellular levels. Restoration of function seems to involve activation of certain compensatory brain regions as a mechanism at the network level to correct the imbalances to the nervous system resulting from loss of dopaminergic neurons in substantia nigra. Clinical studies in China and Korea, in particular, have shown a positive benefit of acupuncture in treating Parkinson's disease, especially in reducing the doses of dopaminergic medications and the associated side effects. However, large and well-controlled clinical trials are still needed to further demonstrate the efficacy and effectiveness of acupuncture in the treatment of Parkinson's disease.展开更多
Cell replacement therapy utilizing mesenchymal stem cells as its main resource holds great promise for ultimate treatment of human neurological disorders.Parkinson＇s disease（PD）is a common,chronic neurodegenerative...Cell replacement therapy utilizing mesenchymal stem cells as its main resource holds great promise for ultimate treatment of human neurological disorders.Parkinson＇s disease（PD）is a common,chronic neurodegenerative disorder hallmarked by localized degeneration of a specific set of dopaminergic neurons within a midbrain sub-region.The specific cell type and confined location of degenerating neurons make cell replacement therapy ideal for PD treatment since it mainly requires replenishment of lost dopaminergic neurons with fresh and functional ones.Endogenous as well as exogenous cell sources have been identified as candidate targets for cell replacement therapy in PD.In this review,umbilical cord mesenchymal stem cells（UCMSCs）are discussed as they provide an inexpensive unlimited reservoir differentiable towards functional dopaminergic neurons that potentially lead to long-lasting behavioral recovery in PD patients.We also present mi RNAs-mediated neuronal differentiation of UCMSCs.The UCMSCs bear a number of outstanding characteristics including their non-tumorigenic,low-immunogenic properties that make them ideal for cell replacement therapy purposes.Nevertheless,more investigations as well as controlled clinical trials are required to thoroughly confirm the efficacy of UCMSCs for therapeutic medical-grade applications in PD.展开更多
BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and fo...BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and folliculogenesis in mice with chemotherapy-induced POF.Human embryonic stem cells(ES)provide an alternative source for mesenchymal stem cells(MSCs)because of their similarities in phenotype and immunomodulatory and anti-inflammatory characteristics.Embryonic stem cell-derived mesenchymal stem cells(ES-MSCs)are attractive candidates for regenerative medicine because of their high proliferation and lack of barriers for harvesting tissue-specific MSCs.However,possible therapeutic effects and underlying mechanisms of transplanted ES-MSCs on cyclophosphamide and busulfan-induced mouse ovarian damage have not been evaluated.AIM To evaluate ES-MSCs vs bone marrow-derived mesenchymal stem cells(BMMSCs)in restoring ovarian function in a mouse model of chemotherapy-induced premature ovarian failure.METHODS Female mice received intraperitoneal injections of different doses of cyclophosphamide and busulfan to induce POF.Either human ES-MSCs or BMMSCs were transplanted into these mice.Ten days after the mice were injected with cyclophosphamide and busulfan and 4 wk after transplantation of the ESMSCs and/or BM-MSCs,we evaluated body weight,estrous cyclicity,folliclestimulating hormone and estradiol hormone concentrations and follicle count were used to evaluate the POF model and cell transplantation.Moreover,terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling,real-time PCR,Western blot analysis and immunohistochemistry and mating was used to evaluate cell transplantation.Enzyme-linked immunosorbent assay was used to analyze vascular endothelial growth factor,insulin-like growth factor 2 and hepatocyte growth factor levels in ES-MSC condition medium in order to investigate the mechanisms that underlie their function.RESULTS The human ES-MSCs significantly restored hormone secretion,survival rate and reproductive function in POF mice,which was similar to the results obtained with BM-MSCs.Gene expression analysis and the terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling assay results indicated that the ES-MSCs and/or BM-MSCs reduced apoptosis in the follicles.Notably,the transplanted mice generated new offspring.The results of different analyses showed increases in antiapoptotic and trophic proteins and genes.CONCLUSION These results suggested that transplantation of human ES-MSCs were similar to BM-MSCs in that they could restore the structure of the injured ovarian tissue and its function in chemotherapy-induced damaged POF mice and rescue fertility.The possible mechanisms of human ES-MSC were related to promotion of follicular development,ovarian secretion,fertility via a paracrine effect and ovarian cell survival.展开更多
Extracellular vesicles(EVs)are nanosized membrane-enclosed compartments that serve as messengers in cell-to-cell communication,both in normal physiology and in pathological conditions.EVs can transfer functional prote...Extracellular vesicles(EVs)are nanosized membrane-enclosed compartments that serve as messengers in cell-to-cell communication,both in normal physiology and in pathological conditions.EVs can transfer functional proteins and genetic information to alter the phenotype and function of recipient cells,which undergo different changes that positively affect their structural and functional integrity.Biological fluids are enriched with several subpopulations of EVs,including exosomes,microvesicles(MVs),and apoptotic bodies carrying several cargoes,such as lipids,proteins,and nucleic acids.EVs associated with the reproductive system are actively involved in the regulation of different physiological events,including gamete maturation,fertilization,and embryo and fetal development.EVs can influence follicle development,oocyte maturation,embryo production,and endometrial-conceptus communication.EVs loaded with cargoes are used to diagnose various diseases,including pregnancy disorders;however,these are dependent on the type of cell of origin and pathological characteristics.EV-derived microRNAs(miRNAs)and proteins in the placenta regulate inflammatory responses and trophoblast invasion through intercellular delivery in the placental microenvironment.This review presents evidence regarding the types of extracellular vesicles,and general aspects of isolation,purification,and characterization of EVs,particularly from various types of embryos.Further,we discuss EVs as mediators and messengers in reproductive biology,the effects of EVs on placentation and pregnancy disorders,the role of EVs in animal reproduction,in the male reproductive system,and mother and embryo cross-communication.In addition,we emphasize the role of microRNAs in embryo implantation and the role of EVs in reproductive and therapeutic medicine.Finally,we discuss the future perspectives of EVs in reproductive biology.展开更多
BACKGROUND Colon cancer cell lines are widely used for research and for the screening of drugs that specifically target the stem cell compartment of colon cancers.It was reported that colon cancer carcinoma specimens ...BACKGROUND Colon cancer cell lines are widely used for research and for the screening of drugs that specifically target the stem cell compartment of colon cancers.It was reported that colon cancer carcinoma specimens contain a subset of leucine-rich repeatcontaining G protein-coupled receptor 5(LGR5)-expressing stem cells,these socalled“tumour-initiating”cells,reminiscent in their properties of the normal intestinal stem cells(ISCs),may explain the apparent heterogeneity of colon cancer cell lines.Also,colon cancer is initiated by aberrant Wnt signaling in ISCs known to express high levels of LGR5.Furthermore,in vivo reports demonstrate the clonal expansion of intestinal adenomas from a single LGR5-expressing cell.AIM To investigate whether colon cancer cell lines contain cancer stem cells and to characterize these putative cancer stem cells.METHODS A portable fluorescent reporter construct based on a conserved fragment of the LGR5 promoter was used to isolate the cell compartments expressing different levels of LGR5 in two widely used colon cancer cell lines(Caco-2 and LoVo).These cells were then characterized according to their proliferation capacity,gene expression signatures of ISC markers,and their tumorigenic properties in vivo and in vitro.RESULTS The data revealed that the LGR5 reporter can be used to identify and isolate a classical intestinal crypt stem cell-like population from the Caco-2,but not from the LoVo,cell lines,in which the cancer stem cell population is more akin to B lymphoma Moloney murine leukemia virus insertion region 1 homolog(+4 crypt)stem cells.This sub-population within Caco-2 cells exhibits an intestinal cancer stem cell gene expression signature and can both self-renew and generate differentiated LGR5 negative progeny.Our data also show that cells expressing high levels of LGR5/enhanced yellow fluorescent protein(EYFP)from this cell line exhibit tumorigenic-like properties in vivo and in vitro.In contrast,cell compartments of LoVo that are expressing high levels of LGR5/EYFP did not show these stem cell-like properties.Thus,cells that exhibit high levels of LGR5/EYFP expression represent the cancer stem cell compartment of Caco-2 colon cancer cells,but not LoVo cells.CONCLUSION Our findings highlight the presence of a spectrum of different ISC-like compartments in different colon cancer cell lines.Their existence is an important consideration for their screening applications and should be taken into account when interpreting drug screening data.We have generated a portable LGR5-reporter that serves as a valuable tool for the identification and isolation of different colon cancer stem cell populations in colon cancer lines.展开更多
Adipose tissue is a compact and well-organized tissue containing a heterogeneous cellular population of progenitor cells,including mesenchymal stromal cells.Due to its availability and accessibility,adipose tissue is ...Adipose tissue is a compact and well-organized tissue containing a heterogeneous cellular population of progenitor cells,including mesenchymal stromal cells.Due to its availability and accessibility,adipose tissue is considered a“stem cell depot.”Adipose tissue products possess anti-inflammatory,anti-fibrotic,antiapoptotic,and immunomodulatory effects.Nanofat,being a compact bundle of stem cells with regenerative and tissue remodeling potential,has potential in translational and regenerative medicine.Considering the wide range of applicability of its reconstructive and regenerative potential,the applications of nanofat can be used in various disciplines.Nanofat behaves on the line of adipose tissuederived mesenchymal stromal cells.At the site of injury,these stromal cells initiate a site-specific reparative response comprised of remodeling of the extracellular matrix,enhanced and sustained angiogenesis,and immune system modulation.These properties of stromal cells provide a platform for the usage of regenerative medicine principles in curbing various diseases.Details about nanofat,including various preparation methods,characterization,delivery methods,evidence on practical applications,and ethical concerns are included in this review.However,appropriate guidelines and preparation protocols for its optimal use in a wide range of clinical applications have yet to be standardized.展开更多
文摘BACKGROUND Knee osteoarthritis(OA)is a degenerative joint disease traditionally viewed through the lens of cartilage degradation.However,emerging evidence positions subchondral bone pathology-particularly bone marrow lesions(BMLs)-as a key contributor to pain,progression,and structural deterioration.Mesenchymal stem cell exhaustion within the osteoarthritic subchondral zone further impairs intrinsic repair mechanisms,reinforcing the rationale for biologic interventions.AIM To evaluate the clinical efficacy of bone marrow aspirate concentrate(BMAC)therapy for knee OA,comparing subchondral vs intra-articular delivery routes,and elucidating the therapeutic impact on symptom relief and structural preservation.METHODS Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines,five clinical studies were included-comprising three randomized controlled trials and two prospective cohorts-with pooled data from 298 knees.Data on functional outcomes,imaging findings,and progression to total knee arthroplasty(TKA)were extracted and qualitatively synthesized.RESULTS Subchondral BMAC injections demonstrated superior improvements compared to intra-articular injection or placebo:Knee Injury and Osteoarthritis Outcome Score improved from 49.1±1.9 to 61.2±6.3 at 12 months(P<0.05),Knee Society Score increased from 57±12 to 87.3±12 at two years,and Western Ontario and McMaster Universities Arthritis Index scores showed significant improvement favoring combined approaches.Magnetic resonance imaging analyses revealed mean BML volume regression of 2.1 cm3,with 80%of knees avoiding TKA over 13-year follow-up.Magnetic resonance imaging analyses revealed regression of BMLs and increased cartilage preservation in subchondral-treated knees.Long-term data indicated delayed progression to TKA and biomechanical improvements(e.g.,Hip-Knee-Ankle angle correction).No major adverse events were reported.CONCLUSION Targeting subchondral bone with BMAC addresses underlying OA pathology and may offer disease-modifying potential beyond symptom relief.These findings support a paradigm shift toward whole-joint biologic therapy,positioning the subchondral matrix as a therapeutic epicenter in OA management.
文摘AIM: To evaluate safety and feasibility of autologous bone marrow-enriched CD34+ hematopoietic stem cell Tx through the hepatic artery in patients with decompensated cirrhosis.METHODS: Four patients with decompensated cirrhosis were included. Approximately 200 mL of the bone marrow of the patients was aspirated, and CD34+ stem cells were selected. Between 3 to 10 million CD34+ cells were isolated. The cells were slowly infused through the hepatic artery of the patients.RESULTS: Patient 1 showed marginal improvement in serum albumin and no significant changes in other test results. In patient 2 prothrombin time was decreased; however, her total bilirubin, serum creatinine, and Model of End-Stage Liver Disease (MELD) score worsened at the end of follow up. In patient 3 there was improvement in serum albumin, porthrombin time (PT), and MELD score. Patient 4 developed radiocontrast nephropathy after the procedure, and progressed to type 1 hepatorenal syndrome and died of liver failure a few days later. Because of the major side effects seen in the last patient, the trial was prematurely stopped.CONCLUSION: Infusion of CD34+ stem cells through the hepatic artery is not safe in decompensated cirrhosis. Radiocontrast nephropathy and hepatorenal syndrome could be major side effects. However, this study doesnot preclude infusion of CD34+ stem cells through other routes.
基金Supported by Science and Technology Program of Shenyang (2009-090063, 2011-F10-222-4-00)
文摘Objective To establish the method of isolation, purification, and identification of human amniotic mesenchymal stem cells (hAMSCs). Methods hAMSCs were isolated from human amniotic membrane by trypsin-collagenase digestion, and cultured in Dulbecco's modified Eagle's medinm/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of immunocytochemistry and flow cytometry. Results The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers CD44, CD90, and vimentin, but not CD45. Conclusions This study establishes a potential method for isolation of hAMSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic characteristics of mesenchymal stem cells.
基金funded by the Research Center for Science and Technology in Medicine(RCSTiM),Tehran University of Medical Sciences,Tehran(TUMS),Tehran,Iran
文摘Several studies have demonstrated that selective serotonin reuptake inhibitor antidepressants can promote neuronal cell proliferation and enhance neuroplasticity both in vitro and in vivo. It is hypothesized that citalopram, a selective serotonin reuptake inhibitor, can promote the neuronal differentiation of adult bone marrow mesenchymal stem cells. Citalopram strongly enhanced neuronal characteristics of the cells derived from bone marrow mesenchymal stem cells. The rate of cell death was decreased in citalopram-treated bone marrow mesenchymal stem cells than in control cells in neurobasal medium. In addition, the cumulative population doubling level of the citalopram-treated cells was signiifcantly increased compared to that of control cells. Also BrdU incorporation was elevated in citalopram-treated cells. These ifndings suggest that citalopram can improve the neuronal-like cell differentiation of bone marrow mesenchymal stem cells by increasing cell proliferation and survival while maintaining their neuronal characteristics.
基金the Scientific and Technological Research Council of Turkey(TÜBİTAK),No.118S738 and No.219S675.
文摘Despite a vast amount of different methods, protocols and cryoprotective agents(CPA), stem cells are often frozen using standard protocols that have beenoptimized for use with cell lines, rather than with stem cells. Relatively fewcomparative studies have been performed to assess the effects of cryopreservationmethods on these stem cells. Dimethyl sulfoxide (DMSO) has been a key agent forthe development of cryobiology and has been used universally for cryopreservation.However, the use of DMSO has been associated with in vitro and in vivotoxicity and has been shown to affect many cellular processes due to changes inDNA methylation and dysregulation of gene expression. Despite studies showingthat DMSO may affect cell characteristics, DMSO remains the CPA of choice, bothin a research setting and in the clinics. However, numerous alternatives to DMSOhave been shown to hold promise for use as a CPA and include albumin,trehalose, sucrose, ethylene glycol, polyethylene glycol and many more. Here, wewill discuss the use, advantages and disadvantages of these CPAs for cryopreservationof different types of stem cells, including hematopoietic stem cells,mesenchymal stromal/stem cells and induced pluripotent stem cells.
基金supported by National Research Programme,“Healthy ageing”(Grant No.SEN-15090)from Research Council of Lithuania
文摘Extracellular vesicles（EVs）provide a novel mechanism of intercellular communication via the transfer of proteins,lipids,and miR NAs between cells.It is now widely accepted that cargo content of EVs depends on cell type and its physiological state.Accordingly,EVs derived from healthy cells may have a comparable therapeutic potential as cells themselves.
基金Supported by the National Natural Science Foundation of China,No.81871568,No.32100643COVID-19 Infection and Prevention Emergency Special Project of Chongqing Education Commission,No.KYYJ202009.
文摘BACKGROUND The immunosuppressive capacity of mesenchymal stem cells(MSCs)is dependent on the“license”of several proinflammatory factors to express immunosuppressive factors such as programmed cell death 1 ligand 1(PD-L1),which determines the clinical therapeutic efficacy of MSCs for inflammatory or immune diseases.In MSCs,interferon-gamma(IFN-γ)is a key inducer of PD-L1 expression,which is synergistically enhanced by tumor necrosis factor-alpha(TNF-α);however,the underlying mechanism is unclear.AIM To reveal the mechanism of pretreated MSCs express high PD-L1 and explore the application of pretreated MSCs in ulcerative colitis.METHODS We assessed PD-L1 expression in human umbilical-cord-derived MSCs(hUC-MSCs)induced by IFN-γand TNF-α,alone or in combination.Additionally,we performed signal pathway inhibitor experiments as well as RNA interference experiments to elucidate the molecular mechanism by which IFN-γalone or in combination with TNF-αinduces PD-L1 expression.Moreover,we used luciferase reporter gene experiments to verify the binding sites of the transcription factors of each signal transduction pathway to the targeted gene promoters.Finally,we evaluated the immunosuppressive capacity of hUC-MSCs treated with IFN-γand TNF-αin both an in vitro mixed lymphocyte culture assay,and in vivo in mice with dextran sulfate sodium-induced acute colitis.RESULTS Our results suggest that IFN-γinduction alone upregulates PD-L1 expression in hUC-MSCs while TNF-αalone does not,and that the co-induction of IFN-γand TNF-αpromotes higher expression of PD-L1.IFN-γinduces hUCMSCs to express PD-L1,in which IFN-γactivates the JAK/STAT1 signaling pathway,up-regulates the expression of the interferon regulatory factor 1(IRF1)transcription factor,promotes the binding of IRF1 and the PD-L1 gene promoter,and finally promotes PD-L1 mRNA.Although TNF-αalone did not induce PD-L1 expression in hUCMSCs,the addition of TNF-αsignificantly enhanced IFN-γ-induced JAK/STAT1/IRF1 activation.TNF-αupregulated IFN-γreceptor expression through activation of the nuclear factor kappa-B signaling pathway,which significantly enhanced IFN-γsignaling.Finally,co-induced hUC-MSCs have a stronger inhibitory effect on lymphocyte proliferation,and significantly ameliorate weight loss,mucosal damage,inflammatory cell infiltration,and up-regulation of inflammatory factors in colitis mice.CONCLUSION Overall,our results suggest that IFN-γand TNF-αenhance both the immunosuppressive ability of hUC-MSCs and their efficacy in ulcerative colitis by synergistically inducing high expression of PD-L1.
基金supported by a Grant-in-Aid for Scientific Research(B)JP21H02808(to TM)JST SPRING JPMJSP2136(to MI)。
文摘Neural stem cells(NSCs)are the source of all neurons and glial cells(astrocytes and oligodendrocytes)in the central nervous system.The adult mammalian brain retains NSCs in the subgranular zone of the dentate gyrus in the hippocampus and ventricular subventricular zone lining the lateral ventricle(Olpe and Jessberger,2022).Adult NSCs in rodents are preserved throughout life and continuously produce new neurons that integrate into the pre-existing neuronal network.
文摘CD34+cells differentiated from mesenchymal stem cells (MSCs) have a strong biological function in cardiovascular regeneration. However, the molecular mechanisms of and the methods to improve the CD34+ cell differentiation from MSCs, especially from human MSCs (hUC-MSCs) are still unclear. In the current study, the effect of CD34 antibody on the CD34+ cell differentiation from human umbilical cord (UC)-derived MSCs (hUC-MSCs) is determined. The results have demonstrated that the expression of cd34 protein is significantly increased in hUC-MSCs treated with CD34 antibody. In addition, the cell proliferation is increased in hUC-MSCs after treatment with CD34 antibody. Moreover, the expression of PI3K, AKT, p-AKT proteins, which are signaling molecules related to stem cell differentiation, is increased by CD34 antibody. The results suggest that CD34 antibody could promote the differentiation of hUC-MSCs into CD34+ cells and PI3K/AKT may be involved in this important process.
文摘Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.
基金financially supported by the Korea Institute of Planning and Evaluation for Technology in Food,Agriculture and Forestry(IPET-RS-2024–00402320)by the Meterials/Parts Technology Development Pro-gram(1415187291,Development of composite formulation with a sustained release(gene)for the treatment of companion animal sarcopenia)funded By the Ministry of Trade,Industry&Energy(MOTIE,Korea)。
文摘Background As the global population increases,the demand for protein sources is expected to increase,driving the demand for cell-based cultivated meat.This study aimed to enhance the productivity of cultivated meat through optimization of the cell source and organization process.Results We engineered fibroblasts into myogenic cells via non-viral introduction of the MYOD1 gene,avoiding viral methods for safety.After confirming the stable derivation of myogenic cells,we combined knockout(KO)of MSTN,a negative regulator of myogenesis,with MYOD1-mediated myogenesis to improve cultivated meat production.Primary cells from MSTN KO cattle exhibited enhanced myogenic potential.Additionally,when tested in immortalized fibroblasts,myostatin treatment reduced MYOD1-induced myogenesis in two-dimensional cultures,while MSTN knockout increased it.To achieve muscle-like cell alignment,we employed digital light processing(DLP)-based three-dimensional(3D)bioprinting to organize cells into 3D groove-shaped hydrogels.These bioactive hydrogels supported stable cell proliferation and significantly improved muscle cell alignment.Upon differentiation into myotubes,the cells demonstrated an ordered alignment,particularly the MSTN KO cells,which showed highly efficient differentiation.Conclusions The integration of genetic modification and advanced DLP 3D bioprinting with groove-patterned hydrogels provides an effective strategy for producing high-quality,muscle-aligned cultivated meat.
文摘“Last scene of all that ends this strange,eventful history,is second childishness and mere oblivion.I am sans teeth,sans eyes,sans taste,sans everything.”William Shakespeare‘As You Like It'Act 2,Sc.7,l.139Aging of the human brain is characterized by a progressive decline of its functional capacity;this decline however varies widely,and cognitive longevity differs substantially between individuals.
基金financially supported by grants from Royan In-stitute(grant No.400000200)Bahar Tashkhis Teb Co.(BTT,9703,9809,and 9903)。
文摘Background: Despite considerable advancements in identifying factors contributing to the development of hepatocellular carcinoma(HCC), the pathogenesis of HCC remains unclear. In many cases, HCC is a consequence of prolonged liver fibrosis, resulting in the formation of an intricate premalignant microenvironment. The accumulation of extracellular matrix(ECM) is a hallmark of premalignant microenvironment. Given the critical role of different matrix components in regulating cell phenotype and function, this study aimed to elucidate the interplay between the fibrotic matrix and malignant features in HCC. Methods: Liver tissues from both control(normal) and carbon tetrachloride(CCl_(4))-induced fibrotic rats were decellularized using sodium dodecyl sulfate(SDS) and Triton X-100. The resulting hydrogel from decellularized ECM was processed into micro-particles via the water-in-oil emulsion method. Microparticles were subsequently incorporated into three-dimensional liver biomimetic micro-tissues(MTs) comprising Huh-7 cells, human umbilical vein endothelial cells(HUVECs), and LX-2 cells. The MTs were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium(MTS) assay at day 11, immunofluorescence staining, immunoblotting, and spheroid migration assay at day 14 after co-culture. Results: Fibrotic matrix from CCl4-treated rat livers significantly enhanced the growth rate of the MTs and their expression of CCND1 as compared to the normal one. Fibrotic matrix, also induced the expression of epithelial-to-mesenchymal transition(EMT)-associated genes such as TWIST1, ACTA2, MMP9, CDH2, and VIMENTIN in the MTs as compared to the normal matrix. Conversely, the expression of CDH1 and hepatic maturation genes HNF4A, ALB, CYP3A4 was decreased in the MTs when the fibrotic matrix was used. Furthermore, the fibrotic matrix increased the migration of the MTs and their secretion of alpha-fetoprotein. Conclusions: Our findings suggest a regulatory role for the fibrotic matrix in promoting cancerous phenotype, which could potentially accelerate the progression of malignancy in the liver.
文摘Since articular cartilage possesses only a weak capac-ity for repair, its regeneration potential is considered one of the most important challenges for orthopedic surgeons. The treatment options, such as marrow stimulation techniques, fail to induce a repair tissue with the same functional and mechanical properties of native hyaline cartilage. Osteochondral transplantation is considered an effective treatment option but is as-sociated with some disadvantages, including donor-site morbidity, tissue supply limitation, unsuitable mechani-cal properties and thickness of the obtained tissue. Although autologous chondrocyte implantation results in reasonable repair, it requires a two-step surgical pro-cedure. Moreover, chondrocytes expanded in culture gradually undergo dedifferentiation, so lose morpho-logical features and specialized functions. In the search for alternative cells, scientists have found mesenchymal stem cells(MSCs) to be an appropriate cellular mate-rial for articular cartilage repair. These cells were origi-nally isolated from bone marrow samples and further investigations have revealed the presence of the cells in many other tissues. Furthermore, chondrogenic dif-ferentiation is an inherent property of MSCs noticedat the time of the cell discovery. MSCs are known to exhibit homing potential to the damaged site at which they differentiate into the tissue cells or secrete a wide spectrum of bioactive factors with regenerative proper-ties. Moreover, these cells possess a considerable im-munomodulatory potential that make them the general donor for therapeutic applications. All of these topics will be discussed in this review.
文摘Acupuncture has been commonly used as an adjuvant therapy or monotherapy in the treatment of Parkinson's disease in China and in other countries.Animal studies have consistently show that this treatment is both neuroprotective, protecting dopaminergic neurons from degeneration and also restorative, restoring tyrosine hydroxylase positive dopaminergic terminals in striatum, resulting in improvements in motor performance in animal models of Parkinsonism. Studies show that this protection is mediated through the same common mechanisms as other neuroprotective agents, including anti-oxidative stress, anti-inflammatory and anti-apoptotic pathways at molecular and cellular levels. Restoration of function seems to involve activation of certain compensatory brain regions as a mechanism at the network level to correct the imbalances to the nervous system resulting from loss of dopaminergic neurons in substantia nigra. Clinical studies in China and Korea, in particular, have shown a positive benefit of acupuncture in treating Parkinson's disease, especially in reducing the doses of dopaminergic medications and the associated side effects. However, large and well-controlled clinical trials are still needed to further demonstrate the efficacy and effectiveness of acupuncture in the treatment of Parkinson's disease.
文摘Cell replacement therapy utilizing mesenchymal stem cells as its main resource holds great promise for ultimate treatment of human neurological disorders.Parkinson＇s disease（PD）is a common,chronic neurodegenerative disorder hallmarked by localized degeneration of a specific set of dopaminergic neurons within a midbrain sub-region.The specific cell type and confined location of degenerating neurons make cell replacement therapy ideal for PD treatment since it mainly requires replenishment of lost dopaminergic neurons with fresh and functional ones.Endogenous as well as exogenous cell sources have been identified as candidate targets for cell replacement therapy in PD.In this review,umbilical cord mesenchymal stem cells（UCMSCs）are discussed as they provide an inexpensive unlimited reservoir differentiable towards functional dopaminergic neurons that potentially lead to long-lasting behavioral recovery in PD patients.We also present mi RNAs-mediated neuronal differentiation of UCMSCs.The UCMSCs bear a number of outstanding characteristics including their non-tumorigenic,low-immunogenic properties that make them ideal for cell replacement therapy purposes.Nevertheless,more investigations as well as controlled clinical trials are required to thoroughly confirm the efficacy of UCMSCs for therapeutic medical-grade applications in PD.
文摘BACKGROUND Premature ovarian failure(POF)affects many adult women less than 40 years of age and leads to infertility.According to previous reports,various tissue-specific stem cells can restore ovarian function and folliculogenesis in mice with chemotherapy-induced POF.Human embryonic stem cells(ES)provide an alternative source for mesenchymal stem cells(MSCs)because of their similarities in phenotype and immunomodulatory and anti-inflammatory characteristics.Embryonic stem cell-derived mesenchymal stem cells(ES-MSCs)are attractive candidates for regenerative medicine because of their high proliferation and lack of barriers for harvesting tissue-specific MSCs.However,possible therapeutic effects and underlying mechanisms of transplanted ES-MSCs on cyclophosphamide and busulfan-induced mouse ovarian damage have not been evaluated.AIM To evaluate ES-MSCs vs bone marrow-derived mesenchymal stem cells(BMMSCs)in restoring ovarian function in a mouse model of chemotherapy-induced premature ovarian failure.METHODS Female mice received intraperitoneal injections of different doses of cyclophosphamide and busulfan to induce POF.Either human ES-MSCs or BMMSCs were transplanted into these mice.Ten days after the mice were injected with cyclophosphamide and busulfan and 4 wk after transplantation of the ESMSCs and/or BM-MSCs,we evaluated body weight,estrous cyclicity,folliclestimulating hormone and estradiol hormone concentrations and follicle count were used to evaluate the POF model and cell transplantation.Moreover,terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling,real-time PCR,Western blot analysis and immunohistochemistry and mating was used to evaluate cell transplantation.Enzyme-linked immunosorbent assay was used to analyze vascular endothelial growth factor,insulin-like growth factor 2 and hepatocyte growth factor levels in ES-MSC condition medium in order to investigate the mechanisms that underlie their function.RESULTS The human ES-MSCs significantly restored hormone secretion,survival rate and reproductive function in POF mice,which was similar to the results obtained with BM-MSCs.Gene expression analysis and the terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling assay results indicated that the ES-MSCs and/or BM-MSCs reduced apoptosis in the follicles.Notably,the transplanted mice generated new offspring.The results of different analyses showed increases in antiapoptotic and trophic proteins and genes.CONCLUSION These results suggested that transplantation of human ES-MSCs were similar to BM-MSCs in that they could restore the structure of the injured ovarian tissue and its function in chemotherapy-induced damaged POF mice and rescue fertility.The possible mechanisms of human ES-MSC were related to promotion of follicular development,ovarian secretion,fertility via a paracrine effect and ovarian cell survival.
文摘Extracellular vesicles(EVs)are nanosized membrane-enclosed compartments that serve as messengers in cell-to-cell communication,both in normal physiology and in pathological conditions.EVs can transfer functional proteins and genetic information to alter the phenotype and function of recipient cells,which undergo different changes that positively affect their structural and functional integrity.Biological fluids are enriched with several subpopulations of EVs,including exosomes,microvesicles(MVs),and apoptotic bodies carrying several cargoes,such as lipids,proteins,and nucleic acids.EVs associated with the reproductive system are actively involved in the regulation of different physiological events,including gamete maturation,fertilization,and embryo and fetal development.EVs can influence follicle development,oocyte maturation,embryo production,and endometrial-conceptus communication.EVs loaded with cargoes are used to diagnose various diseases,including pregnancy disorders;however,these are dependent on the type of cell of origin and pathological characteristics.EV-derived microRNAs(miRNAs)and proteins in the placenta regulate inflammatory responses and trophoblast invasion through intercellular delivery in the placental microenvironment.This review presents evidence regarding the types of extracellular vesicles,and general aspects of isolation,purification,and characterization of EVs,particularly from various types of embryos.Further,we discuss EVs as mediators and messengers in reproductive biology,the effects of EVs on placentation and pregnancy disorders,the role of EVs in animal reproduction,in the male reproductive system,and mother and embryo cross-communication.In addition,we emphasize the role of microRNAs in embryo implantation and the role of EVs in reproductive and therapeutic medicine.Finally,we discuss the future perspectives of EVs in reproductive biology.
基金We thank Professor McGuckin M(MMRI,Brisbane)for providing human colon cancer cell lines(Caco-2,LoVo,and SW480)Dr.Rolfe B(AIBN,Brisbane)for providing mouse NSC-34 cells.
文摘BACKGROUND Colon cancer cell lines are widely used for research and for the screening of drugs that specifically target the stem cell compartment of colon cancers.It was reported that colon cancer carcinoma specimens contain a subset of leucine-rich repeatcontaining G protein-coupled receptor 5(LGR5)-expressing stem cells,these socalled“tumour-initiating”cells,reminiscent in their properties of the normal intestinal stem cells(ISCs),may explain the apparent heterogeneity of colon cancer cell lines.Also,colon cancer is initiated by aberrant Wnt signaling in ISCs known to express high levels of LGR5.Furthermore,in vivo reports demonstrate the clonal expansion of intestinal adenomas from a single LGR5-expressing cell.AIM To investigate whether colon cancer cell lines contain cancer stem cells and to characterize these putative cancer stem cells.METHODS A portable fluorescent reporter construct based on a conserved fragment of the LGR5 promoter was used to isolate the cell compartments expressing different levels of LGR5 in two widely used colon cancer cell lines(Caco-2 and LoVo).These cells were then characterized according to their proliferation capacity,gene expression signatures of ISC markers,and their tumorigenic properties in vivo and in vitro.RESULTS The data revealed that the LGR5 reporter can be used to identify and isolate a classical intestinal crypt stem cell-like population from the Caco-2,but not from the LoVo,cell lines,in which the cancer stem cell population is more akin to B lymphoma Moloney murine leukemia virus insertion region 1 homolog(+4 crypt)stem cells.This sub-population within Caco-2 cells exhibits an intestinal cancer stem cell gene expression signature and can both self-renew and generate differentiated LGR5 negative progeny.Our data also show that cells expressing high levels of LGR5/enhanced yellow fluorescent protein(EYFP)from this cell line exhibit tumorigenic-like properties in vivo and in vitro.In contrast,cell compartments of LoVo that are expressing high levels of LGR5/EYFP did not show these stem cell-like properties.Thus,cells that exhibit high levels of LGR5/EYFP expression represent the cancer stem cell compartment of Caco-2 colon cancer cells,but not LoVo cells.CONCLUSION Our findings highlight the presence of a spectrum of different ISC-like compartments in different colon cancer cell lines.Their existence is an important consideration for their screening applications and should be taken into account when interpreting drug screening data.We have generated a portable LGR5-reporter that serves as a valuable tool for the identification and isolation of different colon cancer stem cell populations in colon cancer lines.
文摘Adipose tissue is a compact and well-organized tissue containing a heterogeneous cellular population of progenitor cells,including mesenchymal stromal cells.Due to its availability and accessibility,adipose tissue is considered a“stem cell depot.”Adipose tissue products possess anti-inflammatory,anti-fibrotic,antiapoptotic,and immunomodulatory effects.Nanofat,being a compact bundle of stem cells with regenerative and tissue remodeling potential,has potential in translational and regenerative medicine.Considering the wide range of applicability of its reconstructive and regenerative potential,the applications of nanofat can be used in various disciplines.Nanofat behaves on the line of adipose tissuederived mesenchymal stromal cells.At the site of injury,these stromal cells initiate a site-specific reparative response comprised of remodeling of the extracellular matrix,enhanced and sustained angiogenesis,and immune system modulation.These properties of stromal cells provide a platform for the usage of regenerative medicine principles in curbing various diseases.Details about nanofat,including various preparation methods,characterization,delivery methods,evidence on practical applications,and ethical concerns are included in this review.However,appropriate guidelines and preparation protocols for its optimal use in a wide range of clinical applications have yet to be standardized.
