Dimethylations of histone H3 lysine 9 and lysine 27 are important epigenetic marks associated with transcription repression. Here, we identified KIAA1718 （KDM7A） as a novel histone demethylase specific for these two...Dimethylations of histone H3 lysine 9 and lysine 27 are important epigenetic marks associated with transcription repression. Here, we identified KIAA1718 （KDM7A） as a novel histone demethylase specific for these two repressing marks. Using mouse embryonic stem cells, we demonstrated that KIAA1718 expression increased at the early phase of neural differentiation. Knockdown of the gene blocked neural differentiation and the effect was rescued by the wild-type human gene, and not by a catalytically inactive mutant. In addition, overexpression of KIAA1718 accelerated neural differentiation. We provide the evidence that the pro-neural differentiation effect of KDM7A is mediated through direct transcriptional activation of FGF4, a signal molecule implicated in neural differentiation. Thus, our study identified a dual-specificity histone demethylase that regulates neural differentiation through FGF4.展开更多
Human Wharton＇s jelly-derived mesenchymal stem cells（h WJ-MSCs）have excellent proliferative ability,differentiation ability,low immunogenicity,and can be easily obtained.However,there are few studies on their appli...Human Wharton＇s jelly-derived mesenchymal stem cells（h WJ-MSCs）have excellent proliferative ability,differentiation ability,low immunogenicity,and can be easily obtained.However,there are few studies on their application in the treatment of ischemic stroke,therefore their therapeutic effect requires further verification.In this study,h WJ-MSCs were transplanted into an ischemic stroke rat model via the tail vein 48 hours after transient middle cerebral artery occlusion.After 4 weeks,neurological functions of the rats implanted with h WJ-MSCs were significantly recovered.Furthermore,many h WJ-MSCs homed to the ischemic frontal cortex whereby they differentiated into neuron-like cells at this region.These results confirm that h WJ-MSCs transplanted into the ischemic stroke rat can differentiate into neuron-like cells to improve rat neurological function and behavior.展开更多
To investigate the pattern of neural differentiation and synaptogenesis in the mouse retina, immunolabeling, Brd U assay and transmission electron microscopy were used. We show that the neuroblastic cell layer is the ...To investigate the pattern of neural differentiation and synaptogenesis in the mouse retina, immunolabeling, Brd U assay and transmission electron microscopy were used. We show that the neuroblastic cell layer is the germinal zone for neural differentiation and retinal lamination. Ganglion cells differentiated initially at embryonic day 13（E13）, and at E18 horizontal cells appeared in the neuroblastic cell layer. Neural stem cells in the outer neuroblastic cell layer differentiated into photoreceptor cells as early as postnatal day 0（P0）, and neural stem cells in the inner neuroblastic cell layer differentiated into bipolar cells at P7. Synapses in the retina were mainly located in the outer and inner plexiform layers. At P7, synaptophysin immunostaining appeared in presynaptic terminals in the outer and inner plexiform layers with button-like structures. After P14, presynaptic buttons were concentrated in outer and inner plexiform layers with strong staining. These data indicate that neural differentiation and synaptogenesis in the retina play important roles in the formation of retinal neural circuitry. Our study showed that the period before P14, especially between P0 and P14, represents a critical period during retinal development. Mouse eye opening occurs during that period, suggesting that cell differentiation and synaptic formation lead to the attainment of visual function.展开更多
Tissue curvature has long been recognized as an important anatomical parameter that affects intracellular behaviors,and there is emerging interest in applying cell-scale curvature as a designer property to drive cell ...Tissue curvature has long been recognized as an important anatomical parameter that affects intracellular behaviors,and there is emerging interest in applying cell-scale curvature as a designer property to drive cell fates for tissue engineering purposes.Although neural cells are known to undergo dramatic and terminal morphological changes during development and curvature-limiting behaviors have been demonstrated in neurite outgrowth studies,there are still crucial gaps in understanding neural cell behaviors,particularly in the context of a three-dimensional(3D)curvature landscape similar to an actual tissue engineering scaffold.In this study,we fabricated two substrates of microcurvature(curvature-substrates)that present a smooth and repeating landscape with focuses of either a concave or a convex pattern.Using these curvature-substrates,we studied the properties of morphological differentiation in N2a neuroblastoma cells.In contrast to other studies where two-dimensional(2D)curvature was demonstrated to limit neurite outgrowth,we found that both the concave and convex substrates acted as continuous and uniform mechanical protrusions that significantly enhanced neural polarity and differentiation with few morphological changes in the main cell body.This enhanced differentiation was manifested in various properties,including increased neurite length,increased nuclear displacement,and upregulation of various neural markers.By demonstrating how the micron-scale curvature landscape induces neuronal polarity,we provide further insights into the design of biomaterials utilizing the influence of surface curvature in neural tissue engineering.展开更多
BACKGROUND Adipose-derived stem cells(ASCs)have been increasingly explored for cell-based medicine because of their numerous advantages in terms of easy availability,high proliferation rate,multipotent differentiation...BACKGROUND Adipose-derived stem cells(ASCs)have been increasingly explored for cell-based medicine because of their numerous advantages in terms of easy availability,high proliferation rate,multipotent differentiation ability and low immunogenicity.In this respect,they have been widely investigated in the last two decades to develop therapeutic strategies for a variety of human pathologies including eye disease.In ocular diseases involving the retina,various cell types may be affected,such as Müller cells,astrocytes,photoreceptors and retinal pigment epithelium(RPE),which plays a fundamental role in the homeostasis of retinal tissue,by secreting a variety of growth factors that support retinal cells.AIM To test ASC neural differentiation using conditioned medium(CM)from an RPE cell line(ARPE-19).METHODS ASCs were isolated from adipose tissue,harvested from the subcutaneous region of healthy donors undergoing liposuction procedures.Four ASC culture conditions were investigated:ASCs cultured in basal Dulbecco's Modified Eagle Medium(DMEM);ASCs cultured in serum-free DMEM;ASCs cultured in serumfree DMEM/F12;and ASCs cultured in a CM from ARPE-19,a spontaneously arising cell line with a normal karyotype derived from a human RPE.Cell proliferation rate and viability were assessed by crystal violet and MTT assays at 1,4and 8 d of culture.At the same time points,ASC neural differentiation was evaluated by immunocytochemistry and western blot analysis for typical neuronal and glial markers:Nestin,neuronal specific enolase(NSE),protein gene product(PGP)9.5,and glial fibrillary acidic protein(GFAP).RESULTS Depending on the culture medium,ASC proliferation rate and viability showed some significant differences.Overall,less dense populations were observed in serum-free cultures,except for ASCs cultured in ARPE-19 serum-free CM.Moreover,a different cell morphology was seen in these cultures after 8 d of treatment,with more elongated cells,often showing cytoplasmic ramifications.Immunofluorescence results and western blot analysis were indicative of ASC neural differentiation.In fact,basal levels of neural markers detected under control conditions significantly increased when cells were cultured in ARPE-19 CM.Specifically,neural marker overexpression was more marked at 8 d.The most evident increase was observed for NSE and GFAP,a modest increase was observed for nestin,and less relevant changes were observed for PGP9.5.CONCLUSION The presence of growth factors produced by ARPE-19 cells in tissue culture induces ASCs to express neural differentiation markers typical of the neuronal and glial cells of the retina.展开更多
Targeted stem cell delivery utilizing a magnetic actuation system is an emerging technology in stem cell engineering that efficiently targets stem cells in specific areas in vitro.However,integrating precise magnetic ...Targeted stem cell delivery utilizing a magnetic actuation system is an emerging technology in stem cell engineering that efficiently targets stem cells in specific areas in vitro.However,integrating precise magnetic control systems with selective neural differentiation has not yet been widely considered for building successful neural networks.Challenges arise in creating targeted functional neuronal networks,largely due to difficulties in simultaneously controlling the positions of stem cells and selectively stimulating their differentiation.These challenges often result in suboptimal differentiation rates and abnormalities in transplanted neural stem cells.In contrast,ultrasound stimulation has superior tissue penetration and focusing capability,and represents a promising noninvasive neural stimulation technique capable of modulating neural activity and promoting selective differentiation into neuronal stem cells.In this study,we introduce a method for targeted neural differentiation using localized ultrasonic stimulation with a piezoelectric micromachined ultrasound transducer(pMUT)array.Differentiation was assessed quantitatively by monitoring neurite outgrowth as the ultrasound intensity was increased.The neurite length of cells ultrasonically stimulated for 40 min was found to have increased,compared to the non-stimulated group(119.9±34.3μm vs.63.2±17.3μm,respectively).Targeted differentiation was confirmed by measuring neurite lengths,where selective ultrasound stimulation induced differentiation in cells that were precisely delivered via an electromagnetic system.Magnetic cell-based robots reaching the area of localized ultrasound stimulation were confirmed to have enhanced differentiation.This research demonstrated the potential of the combination of precise stem cell delivery with selective neural differentiation to establish functional neural networks.展开更多
While it is known that mechanical dynamics are influential in neural differentiation for critical processes like neurogenesis or neurodegeneration, studies on neural stem cell therapies usually focus on biochemical in...While it is known that mechanical dynamics are influential in neural differentiation for critical processes like neurogenesis or neurodegeneration, studies on neural stem cell therapies usually focus on biochemical interactions rather than mechanical aspects, frequently resulting in low efficacy and unfulfilled potential. Therefore, current studies are attempting to elucidate the effect of mechanical stimulus on neural performance using conventional two-dimensional(2D) planar substrates. Yet, these2D substrates fail to capture the defining three-dimensional(3D) characteristics of the in vivo neural stem cell environment.To complete this research gap, we synthesized a series of soft and elastic 3D hydrogels to mimic the neural tissue mechanical environment for 3D cell culture, using long-chain polyethylene glycol diacrylate(PEGDA) and gelatin-methacryloyl(Gel MA).By varying the concentration of the polymer, we obtained biomimicking hydrogels with a tensile modulus as low as 10 k Pa and a compressive modulus as low as 0.8 k Pa. The in vitro results demonstrated that Gel MA-PEGDA hydrogels have the high biocompatibility required to support neural cell growth, proliferation, and differentiation, as well as neurite outgrowth. We then studied the effect of mechanical stretching on the behaviors of neural cells and observed that mechanical stretching could significantly enhance neurite extension and axon elongation. In addition, the neurites were more directionally oriented to the stretching direction. Immunocytochemistry and relative gene expression data also suggested that mechanical tension could upregulate the expression of neural differentiation protein and genes, including GFAP and βIII-Tubulin. Overall, this study shows that in addition to the specific mechanical properties of Gel MA-PEGDA that improve neural differentiation towards specific lineages, hydrogel stretching is also a potentially attractive strategy to improve the therapeutic outcomes of neural stem cell therapies.展开更多
The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model ...The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model for dissecting human development because of unlimited self-renewal and the capacity to differentiate into nearly all cell types in the human body. In this study,using a chemical defined neural induction protocol and molecular profiling, we identified Fez family zinc finger 1 (FEZF1) as a potential regulator of early human neural development. FEZF1 is rapidly up-regulated during neural differentiation in hESCs and expressed before PAX6, a well-established marker of early human neural induction. We generated FEZF1-knockout H1 hESC lines using CRISPR-CAS9 technology and found that depletion of FEZF1 abrogates neural differentiation of hESCs. Moreover,loss of FEZF1 impairs the pluripotency exit of hESCs during neural specification, which partially explains the neural induction defect caused by FEZF1 deletion. However, enforced expression of FEZF1 itself fails to drive neural differentiation in hESCs,suggesting that FEZF1 is necessary but not sufficient for neural differentiation from hESCs. Taken together, our findings identify one of the earliest regulators expressed upon neural induction and provide insight into early neural development in human.展开更多
OBJECTIVE： To summarize the biological characteristics of neural stem cells, and the separation, purification. differentiation and source of neural stem cells. DATA SOURCES ： An online search of Pubmed database was ...OBJECTIVE： To summarize the biological characteristics of neural stem cells, and the separation, purification. differentiation and source of neural stem cells. DATA SOURCES ： An online search of Pubmed database was undertaken to identify English articles about the growth of neural stem cells in vitro published from January 2000 to October 2006 by using the keywords of ＂neural stem cells, bone marrow mesenchymal stem cells （BMSCs）, umbilical cord blood stem cells, embryonic stem cells （ESC）, separation methods, neural growth factor＂. And relevant articles published in IEEE/IEE Electronic Library （IEL） database, Springer Link database and Kluwer Online Journals were also searched, Chinese relevant articles published between January 2000 to October 2006 were searched with the same keywords in Chinese in Chinese journal full-text database. STUDY SELECTION ： The articles were primarily screened, and then the full-texts were searched. Inclusive criteria： （1） Articles relevant to the biological characteristics and classification of neural stem cells; （2） Articles about the source, separation and differentiation of the ESCs, BMSCs and umbilical cord blood stem cells. The repetitive studies and reviews were excluded. DATA EXTRACTION ： Thirty articles were selected from 203 relevant articles according to the inclusive criteria Articles were excluded because of repetition and reviews. DATA SYNTHESES ： Neural stem cells have the ability of self-renewing and high differentiation, and they are obtained from ESCs, nerve tissue, nerve system, BMSCs and umbilical cord blood stem cells. ESCs can be separated by means of mechanical dissociation is better than that of the trypsin digestion, BMSCs by density gradient centrifuge separation, hemolysis, whole-blood culture, etc., and umbilical cord blood stem ceils by Ficoil density gradient centrifugation, hydroxyethyl starch （HES） centrifugation sedimentation, etc. Neural growth factor （NGF） and other factors play an important role in the growth of NSCs, such as transforming growth factor （TGF） is an important player in repairing organs, NGF accelerates the process of growth, insulin-like growth factor serves importantly in the differentiation of stem cells into neuron-like cells. CONCLUSION ： As unipotent stem cells, NSCs have the abilities of self-renewal and potential of high differentiation. The method of mechanical dissociation is better than trypsin digestion in e separating ESCs. However, density gradient centrifuge separation is better than other methods in the separation of the BMSCs. NGF and other factors play an important role in the growth of NSCs.展开更多
Biological studies typically rely on a simple monolayer cell culture,which does not reflect the complex functional characteristics of human tissues and organs,or their real response to external stimuli.Microfluidic te...Biological studies typically rely on a simple monolayer cell culture,which does not reflect the complex functional characteristics of human tissues and organs,or their real response to external stimuli.Microfluidic technology has advantages of high-throughput screening,accurate control of the fluid velocity,low cell consumption,long-term culture,and high integration.By combining the multipotential differentiation of neural stem cells with high throughput and the integrated characteristics of microfluidic technology,an in vitro model of a functionalized neurovascular unit was established using human neural stem cell-derived neurons,astrocytes,oligodendrocytes,and a functional microvascular barrier.The model comprises a multi-layer vertical neural module and vascular module,both of which were connected with a syringe pump.This provides controllable conditions for cell inoculation and nutrient supply,and simultaneously simulates the process of ischemic/hypoxic injury and the process of inflammatory factors in the circulatory system passing through the blood-brain barrier and then acting on the nerve tissue in the brain.The in vitro functionalized neurovascular unit model will be conducive to central nervous system disease research,drug screening,and new drug development.展开更多
Fimbria-fornix transection induces both exogenous and endogenous neural stem cells to differentiate into neurons in the hippocampus.This indicates that the denervated hippocampus provides an environment for neuronal d...Fimbria-fornix transection induces both exogenous and endogenous neural stem cells to differentiate into neurons in the hippocampus.This indicates that the denervated hippocampus provides an environment for neuronal differentiation of neural stem cells.However,the pathways and mechanisms in this process are still unclear.Seven days after fimbria fornix transection,our reverse transcription polymerase chain reaction,western blot assay,and enzyme linked immunosorbent assay results show a significant increase in ciliary neurotrophic factor m RNA and protein expression in the denervated hippocampus.Moreover,neural stem cells derived from hippocampi of fetal（embryonic day 17） Sprague-Dawley rats were treated with ciliary neurotrophic factor for 7 days,with an increased number of microtubule associated protein-2-positive cells and decreased number of glial fibrillary acidic protein-positive cells detected.Our results show that ciliary neurotrophic factor expression is up-regulated in the denervated hippocampus,which may promote neuronal differentiation of neural stem cells in the denervated hippocampus.展开更多
Purinergic receptors are among the first cell surface receptors expressed during embryonic development （Burnstock and Ulrich, 2011）. These are characterized based on their pharmacological properties of being activat...Purinergic receptors are among the first cell surface receptors expressed during embryonic development （Burnstock and Ulrich, 2011）. These are characterized based on their pharmacological properties of being activated by adenosine or purine/pyrimidine nucleotides as P1 and P2 receptors. P2 receptors are further classified by their structure as P2Y metabotropic and P2X ionotropic receptors.展开更多
Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural dif...Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.展开更多
Many blinding diseases,such as retinitis pigmentosa,age-related macular degeneration,and glaucoma involve the permanent loss of retinal neurons,especially photoreceptors or the centrally projecting retinal ganglion ce...Many blinding diseases,such as retinitis pigmentosa,age-related macular degeneration,and glaucoma involve the permanent loss of retinal neurons,especially photoreceptors or the centrally projecting retinal ganglion cells.Stem cells have been proposed as a potential source of cells for neuronal transplantation.展开更多
Previous in vivo experiments have shown that human umbilical cord blood mesenchymal stem cells can promote the proliferation and differentiation of damaged celts, and help to repair damaged sites, Recent studies have ...Previous in vivo experiments have shown that human umbilical cord blood mesenchymal stem cells can promote the proliferation and differentiation of damaged celts, and help to repair damaged sites, Recent studies have reported that umbilical cord blood-derived mesenchymal stem cells can differentiate into neurons and glial cells. Recent studies have reported that the repair mechanisms underlying cord blood stern cells involve the replacement of damaged cells and mediation of the local micro-environment.展开更多
A microgravity environment has been shown to cause ocular damage and affect visual acuity,but the underlying mechanisms remain unclear.Therefore,we established an animal model of weightlessness via tail suspension to ...A microgravity environment has been shown to cause ocular damage and affect visual acuity,but the underlying mechanisms remain unclear.Therefore,we established an animal model of weightlessness via tail suspension to examine the pathological changes and molecular mechanisms of retinal damage under microgravity.After 4 weeks of tail suspension,there were no notable alterations in retinal function and morphology,while after 8 weeks of tail suspension,significant reductions in retinal function were observed,and the outer nuclear layer was thinner,with abundant apoptotic cells.To investigate the mechanism underlying the degenerative changes that occurred in the outer nuclear layer of the retina,proteomics was used to analyze differentially expressed proteins in rat retinas after 8 weeks of tail suspension.The results showed that the expression levels of fibroblast growth factor 2(also known as basic fibroblast growth factor)and glial fibrillary acidic protein,which are closely related to Müller cell activation,were significantly upregulated.In addition,Müller cell regeneration and Müller cell gliosis were observed after 4 and 8 weeks,respectively,of simulated weightlessness.These findings indicate that Müller cells play an important regulatory role in retinal outer nuclear layer degeneration during weightlessness.展开更多
Neuroinflammation is the primary driver and signature of many neurodevelopmental disorders.However,because neurodevelopmental disorders caused by neuroinflammation are difficult to detect at the early stage,their prog...Neuroinflammation is the primary driver and signature of many neurodevelopmental disorders.However,because neurodevelopmental disorders caused by neuroinflammation are difficult to detect at the early stage,their progression remains unclear.To date,neither animal experiments nor in vitro models have uncovered their early developmental characteristics caused by neuroinflammation.In this study,we developed a neurovascular-unit-on-a-chip(NVU-on-a-chip)to model inflammation-induced neurodevelopmental disorders.With the chip,dynamic visualization of the progression caused by neuroinflammation was clearly demonstrated,and the changes in angiogenesis and neural differentiation under neuroinflammation were replicated.In addition,the activation of astrocytes and damage to neurons and capillaries at the early stage of neurodevelopmental disorders were observed.The results revealed for the first time the structural disruption of the neurovascular units and the neurovascular coupling failure caused by neuroinflammation.Furthermore,the outcomes of anti-inflammatory intervention using ibuprofen were preliminarily demonstrated.This work provides insights into the early progression of neurodevelopmental disorders caused by neuroinflammation and offers a platform for the development of therapeutic strategies for neuroinflammation.展开更多
The mechanisms of the transplantation of neural stem cells(NSCs)in the treatment of Alzheimer’s disease remain poorly understood.In this study,NSCs were transplanted into the hippocampal CA1 region of the rTg(tau P30...The mechanisms of the transplantation of neural stem cells(NSCs)in the treatment of Alzheimer’s disease remain poorly understood.In this study,NSCs were transplanted into the hippocampal CA1 region of the rTg(tau P301L)4510 mouse model,a tauopathy model that is thought to reflect the tau pathology associated with Alzheimer’s disease.The results revealed that NSC transplantation reduced the abnormal aggregation of tau,resulting in significant improvements in the short-term memory of the tauopathy model mice.Compared with wild-type and phosphate-buffered saline(PBS)-treated mice,mice that received NSC transplantations were characterized by changes in the expression of multiple proteins in brain tissue,particularly those related to the regulation of tau aggregation or misfolding.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis and Gene Ontology(GO)function analysis revealed that these proteins were primarily enriched in pathways associated with long-term potentiation,neurogenesis,and other neurobiological processes.Changes in the expression levels of key proteins were verified by western blot assays.These data provided clues to improve the understanding of the functional capacity associated with NSC transplantation in Alzheimer’s disease treatment.This study was approved by the Beijing Animal Ethics Association and Ethics Committee of Beijing Institute of Technology(approval No.SYXK-BIT-school of life science-2017-M03)in 2017.展开更多
Previous studies have shown that caveolin-1 is involved in regulating the differentiation of mesenchymal stem cells.However,its role in the differentiation of human adipose mesenchymal stem cells into dopaminergic neu...Previous studies have shown that caveolin-1 is involved in regulating the differentiation of mesenchymal stem cells.However,its role in the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons remains unclear.The aim of this study was to investigate whether caveolin-1 regulates the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.We also examined whether the expression of caveolin-1 could be modulated by RNA interference technology to promote the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.The differentiation of human adipose mesenchymal stem cells into dopaminergic neurons was evaluated morphologically and by examining expression of the markers tyrosine hydroxylase,Lmx1a and Nurr1.The analyses revealed that during the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons,the expression of caveolin-1 is decreased.Notably,the downregulation of caveolin-1 promoted the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons,and it increased the expression of tyrosine hydroxylase,Lmx1a and Nurr1.Together,our findings suggest that caveolin-1 plays a negative regulatory role in the differentiation of dopaminergic-like neurons from stem cells,and it may therefore be a potential molecular target for strategies for regulating the differentiation of these cells.This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Dalian Medical University of China(approval No.PJ-KS-KY-2020-54)on March 7,2017.展开更多
The difference between Noggin and basic fibroblast growth factor for the neural precursor differen- tiation from human embryonic stem cells has not been studied. In this study, 100 tJg/L Noggin or 20 IJg/L basic fibro...The difference between Noggin and basic fibroblast growth factor for the neural precursor differen- tiation from human embryonic stem cells has not been studied. In this study, 100 tJg/L Noggin or 20 IJg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen- tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro- scope. Immunofluorescence staining revealed expression levels of Nestin, [3-111 Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in- creases the differentiation of neural precursors.展开更多
基金Supplementary information is linked to the online version of the paper on the Cell Research website.Acknowledgments We thank Anning Lin （The University of Chicago） for the critical reading of the paper, members in the Chen lab for technical help, the cell biology and molecular biology core facilities for confocal study and Q-PCR, and Shanghai Biochip Co Ltd. for microarray analysis. The H3K27me2 antibody was kindly provided by Li Tang （Fudan University） and Thomas Jenuwein （Research Institute of Molecular Pathology, The Vienna Biocenter）. This work was supported by the National Basic Research Program of China （2007CB957900, 2006CB943902, 2007CB947101, 2008KR0695, 2009CB941100, 2005CB522704）, the Chinese Academy of Sciences （KSCX2-YW-R-04）, the National Natural Science Foundation of China （90919026, 30870538,30623003, 30721065, 30830034, 90919046）, the Shanghai Pujiang Program （0757S11361）, the Shanghai Key Project of Basic Science Research （06DJ14001, 06DZ22032, 08DJ1400501）, and the Council of Shanghai Municipal Government for Science and Technology （088014199）.
文摘Dimethylations of histone H3 lysine 9 and lysine 27 are important epigenetic marks associated with transcription repression. Here, we identified KIAA1718 （KDM7A） as a novel histone demethylase specific for these two repressing marks. Using mouse embryonic stem cells, we demonstrated that KIAA1718 expression increased at the early phase of neural differentiation. Knockdown of the gene blocked neural differentiation and the effect was rescued by the wild-type human gene, and not by a catalytically inactive mutant. In addition, overexpression of KIAA1718 accelerated neural differentiation. We provide the evidence that the pro-neural differentiation effect of KDM7A is mediated through direct transcriptional activation of FGF4, a signal molecule implicated in neural differentiation. Thus, our study identified a dual-specificity histone demethylase that regulates neural differentiation through FGF4.
基金supported by the National Natural Science Foundation of China,No.31171038the Natural Science Foundation of Jiangsu Province of China,No.BK2011385+3 种基金the "333" Program Funding of Jiangsu Province of China,No.BRA2016450the Training Program of Innovation and Entrepreneurship for Undergraduates of Nantong University of China,No.201510304033Z,201610304053Zthe Training Program of Innovation and Entrepreneurship for Graduates of Nantong University of China,No.YKC14050,YKC15046a grant from Funds for the Priority Academic Program Development of Jiangsu Higher Education Institutions of China
文摘Human Wharton＇s jelly-derived mesenchymal stem cells（h WJ-MSCs）have excellent proliferative ability,differentiation ability,low immunogenicity,and can be easily obtained.However,there are few studies on their application in the treatment of ischemic stroke,therefore their therapeutic effect requires further verification.In this study,h WJ-MSCs were transplanted into an ischemic stroke rat model via the tail vein 48 hours after transient middle cerebral artery occlusion.After 4 weeks,neurological functions of the rats implanted with h WJ-MSCs were significantly recovered.Furthermore,many h WJ-MSCs homed to the ischemic frontal cortex whereby they differentiated into neuron-like cells at this region.These results confirm that h WJ-MSCs transplanted into the ischemic stroke rat can differentiate into neuron-like cells to improve rat neurological function and behavior.
基金supported by the National Natural Science Foundation of China,No.31070952 and U1204311the Scientific Research Foundation of Henan University of China,No.0000A40475 and 0000A40356
文摘To investigate the pattern of neural differentiation and synaptogenesis in the mouse retina, immunolabeling, Brd U assay and transmission electron microscopy were used. We show that the neuroblastic cell layer is the germinal zone for neural differentiation and retinal lamination. Ganglion cells differentiated initially at embryonic day 13（E13）, and at E18 horizontal cells appeared in the neuroblastic cell layer. Neural stem cells in the outer neuroblastic cell layer differentiated into photoreceptor cells as early as postnatal day 0（P0）, and neural stem cells in the inner neuroblastic cell layer differentiated into bipolar cells at P7. Synapses in the retina were mainly located in the outer and inner plexiform layers. At P7, synaptophysin immunostaining appeared in presynaptic terminals in the outer and inner plexiform layers with button-like structures. After P14, presynaptic buttons were concentrated in outer and inner plexiform layers with strong staining. These data indicate that neural differentiation and synaptogenesis in the retina play important roles in the formation of retinal neural circuitry. Our study showed that the period before P14, especially between P0 and P14, represents a critical period during retinal development. Mouse eye opening occurs during that period, suggesting that cell differentiation and synaptic formation lead to the attainment of visual function.
基金supported by the Inter-Departmental Open Project of State Key Laboratory in Ultra-Precision Machining Technology(SKL-UPMT,No.P0033576).
文摘Tissue curvature has long been recognized as an important anatomical parameter that affects intracellular behaviors,and there is emerging interest in applying cell-scale curvature as a designer property to drive cell fates for tissue engineering purposes.Although neural cells are known to undergo dramatic and terminal morphological changes during development and curvature-limiting behaviors have been demonstrated in neurite outgrowth studies,there are still crucial gaps in understanding neural cell behaviors,particularly in the context of a three-dimensional(3D)curvature landscape similar to an actual tissue engineering scaffold.In this study,we fabricated two substrates of microcurvature(curvature-substrates)that present a smooth and repeating landscape with focuses of either a concave or a convex pattern.Using these curvature-substrates,we studied the properties of morphological differentiation in N2a neuroblastoma cells.In contrast to other studies where two-dimensional(2D)curvature was demonstrated to limit neurite outgrowth,we found that both the concave and convex substrates acted as continuous and uniform mechanical protrusions that significantly enhanced neural polarity and differentiation with few morphological changes in the main cell body.This enhanced differentiation was manifested in various properties,including increased neurite length,increased nuclear displacement,and upregulation of various neural markers.By demonstrating how the micron-scale curvature landscape induces neuronal polarity,we provide further insights into the design of biomaterials utilizing the influence of surface curvature in neural tissue engineering.
基金Supported by University of Catania,Italy,“Piano Triennale per la Ricerca 2020-2022–Grant PIACERI,project“NanoRet””.
文摘BACKGROUND Adipose-derived stem cells(ASCs)have been increasingly explored for cell-based medicine because of their numerous advantages in terms of easy availability,high proliferation rate,multipotent differentiation ability and low immunogenicity.In this respect,they have been widely investigated in the last two decades to develop therapeutic strategies for a variety of human pathologies including eye disease.In ocular diseases involving the retina,various cell types may be affected,such as Müller cells,astrocytes,photoreceptors and retinal pigment epithelium(RPE),which plays a fundamental role in the homeostasis of retinal tissue,by secreting a variety of growth factors that support retinal cells.AIM To test ASC neural differentiation using conditioned medium(CM)from an RPE cell line(ARPE-19).METHODS ASCs were isolated from adipose tissue,harvested from the subcutaneous region of healthy donors undergoing liposuction procedures.Four ASC culture conditions were investigated:ASCs cultured in basal Dulbecco's Modified Eagle Medium(DMEM);ASCs cultured in serum-free DMEM;ASCs cultured in serumfree DMEM/F12;and ASCs cultured in a CM from ARPE-19,a spontaneously arising cell line with a normal karyotype derived from a human RPE.Cell proliferation rate and viability were assessed by crystal violet and MTT assays at 1,4and 8 d of culture.At the same time points,ASC neural differentiation was evaluated by immunocytochemistry and western blot analysis for typical neuronal and glial markers:Nestin,neuronal specific enolase(NSE),protein gene product(PGP)9.5,and glial fibrillary acidic protein(GFAP).RESULTS Depending on the culture medium,ASC proliferation rate and viability showed some significant differences.Overall,less dense populations were observed in serum-free cultures,except for ASCs cultured in ARPE-19 serum-free CM.Moreover,a different cell morphology was seen in these cultures after 8 d of treatment,with more elongated cells,often showing cytoplasmic ramifications.Immunofluorescence results and western blot analysis were indicative of ASC neural differentiation.In fact,basal levels of neural markers detected under control conditions significantly increased when cells were cultured in ARPE-19 CM.Specifically,neural marker overexpression was more marked at 8 d.The most evident increase was observed for NSE and GFAP,a modest increase was observed for nestin,and less relevant changes were observed for PGP9.5.CONCLUSION The presence of growth factors produced by ARPE-19 cells in tissue culture induces ASCs to express neural differentiation markers typical of the neuronal and glial cells of the retina.
基金financially supported by the National Convergence Research of Scientific Challenges through the National Research Foundation of Korea(NRF)(no.2021M3F7A1082275)funded by the Ministry of Science and ICT.
文摘Targeted stem cell delivery utilizing a magnetic actuation system is an emerging technology in stem cell engineering that efficiently targets stem cells in specific areas in vitro.However,integrating precise magnetic control systems with selective neural differentiation has not yet been widely considered for building successful neural networks.Challenges arise in creating targeted functional neuronal networks,largely due to difficulties in simultaneously controlling the positions of stem cells and selectively stimulating their differentiation.These challenges often result in suboptimal differentiation rates and abnormalities in transplanted neural stem cells.In contrast,ultrasound stimulation has superior tissue penetration and focusing capability,and represents a promising noninvasive neural stimulation technique capable of modulating neural activity and promoting selective differentiation into neuronal stem cells.In this study,we introduce a method for targeted neural differentiation using localized ultrasonic stimulation with a piezoelectric micromachined ultrasound transducer(pMUT)array.Differentiation was assessed quantitatively by monitoring neurite outgrowth as the ultrasound intensity was increased.The neurite length of cells ultrasonically stimulated for 40 min was found to have increased,compared to the non-stimulated group(119.9±34.3μm vs.63.2±17.3μm,respectively).Targeted differentiation was confirmed by measuring neurite lengths,where selective ultrasound stimulation induced differentiation in cells that were precisely delivered via an electromagnetic system.Magnetic cell-based robots reaching the area of localized ultrasound stimulation were confirmed to have enhanced differentiation.This research demonstrated the potential of the combination of precise stem cell delivery with selective neural differentiation to establish functional neural networks.
基金supported by the Youth Program of National Natural Science Foundation of China (No.11702233)。
文摘While it is known that mechanical dynamics are influential in neural differentiation for critical processes like neurogenesis or neurodegeneration, studies on neural stem cell therapies usually focus on biochemical interactions rather than mechanical aspects, frequently resulting in low efficacy and unfulfilled potential. Therefore, current studies are attempting to elucidate the effect of mechanical stimulus on neural performance using conventional two-dimensional(2D) planar substrates. Yet, these2D substrates fail to capture the defining three-dimensional(3D) characteristics of the in vivo neural stem cell environment.To complete this research gap, we synthesized a series of soft and elastic 3D hydrogels to mimic the neural tissue mechanical environment for 3D cell culture, using long-chain polyethylene glycol diacrylate(PEGDA) and gelatin-methacryloyl(Gel MA).By varying the concentration of the polymer, we obtained biomimicking hydrogels with a tensile modulus as low as 10 k Pa and a compressive modulus as low as 0.8 k Pa. The in vitro results demonstrated that Gel MA-PEGDA hydrogels have the high biocompatibility required to support neural cell growth, proliferation, and differentiation, as well as neurite outgrowth. We then studied the effect of mechanical stretching on the behaviors of neural cells and observed that mechanical stretching could significantly enhance neurite extension and axon elongation. In addition, the neurites were more directionally oriented to the stretching direction. Immunocytochemistry and relative gene expression data also suggested that mechanical tension could upregulate the expression of neural differentiation protein and genes, including GFAP and βIII-Tubulin. Overall, this study shows that in addition to the specific mechanical properties of Gel MA-PEGDA that improve neural differentiation towards specific lineages, hydrogel stretching is also a potentially attractive strategy to improve the therapeutic outcomes of neural stem cell therapies.
基金supported by National Basic Research Program of China (2015CB964902 to Jiaxi Zhou and SQ2016ZY05002105 to Hongtao Wang)CAMS Initiative for Innovative Medicine (2016-I2M-1-018, 2016-I2M-3-002)+2 种基金National Natural Science Foundation of China (81530008, 31671541 to Jiaxi Zhou, 31500949 to Hongtao Wang)Tianjin Natural Science Foundation (16JCZDJC33100 to Jiaxi Zhou)3332015128 supported by PUMC Youth Fund and Fundamental Research Funds for the Central Universities to Dr. Hongtao Wang
文摘The understanding of the mechanism underlying human neural development has been hampered due to lack of a cellular system and complicated ethical issues. Human embryonic stem cells (hESCs) provide an invaluable model for dissecting human development because of unlimited self-renewal and the capacity to differentiate into nearly all cell types in the human body. In this study,using a chemical defined neural induction protocol and molecular profiling, we identified Fez family zinc finger 1 (FEZF1) as a potential regulator of early human neural development. FEZF1 is rapidly up-regulated during neural differentiation in hESCs and expressed before PAX6, a well-established marker of early human neural induction. We generated FEZF1-knockout H1 hESC lines using CRISPR-CAS9 technology and found that depletion of FEZF1 abrogates neural differentiation of hESCs. Moreover,loss of FEZF1 impairs the pluripotency exit of hESCs during neural specification, which partially explains the neural induction defect caused by FEZF1 deletion. However, enforced expression of FEZF1 itself fails to drive neural differentiation in hESCs,suggesting that FEZF1 is necessary but not sufficient for neural differentiation from hESCs. Taken together, our findings identify one of the earliest regulators expressed upon neural induction and provide insight into early neural development in human.
文摘OBJECTIVE： To summarize the biological characteristics of neural stem cells, and the separation, purification. differentiation and source of neural stem cells. DATA SOURCES ： An online search of Pubmed database was undertaken to identify English articles about the growth of neural stem cells in vitro published from January 2000 to October 2006 by using the keywords of ＂neural stem cells, bone marrow mesenchymal stem cells （BMSCs）, umbilical cord blood stem cells, embryonic stem cells （ESC）, separation methods, neural growth factor＂. And relevant articles published in IEEE/IEE Electronic Library （IEL） database, Springer Link database and Kluwer Online Journals were also searched, Chinese relevant articles published between January 2000 to October 2006 were searched with the same keywords in Chinese in Chinese journal full-text database. STUDY SELECTION ： The articles were primarily screened, and then the full-texts were searched. Inclusive criteria： （1） Articles relevant to the biological characteristics and classification of neural stem cells; （2） Articles about the source, separation and differentiation of the ESCs, BMSCs and umbilical cord blood stem cells. The repetitive studies and reviews were excluded. DATA EXTRACTION ： Thirty articles were selected from 203 relevant articles according to the inclusive criteria Articles were excluded because of repetition and reviews. DATA SYNTHESES ： Neural stem cells have the ability of self-renewing and high differentiation, and they are obtained from ESCs, nerve tissue, nerve system, BMSCs and umbilical cord blood stem cells. ESCs can be separated by means of mechanical dissociation is better than that of the trypsin digestion, BMSCs by density gradient centrifuge separation, hemolysis, whole-blood culture, etc., and umbilical cord blood stem ceils by Ficoil density gradient centrifugation, hydroxyethyl starch （HES） centrifugation sedimentation, etc. Neural growth factor （NGF） and other factors play an important role in the growth of NSCs, such as transforming growth factor （TGF） is an important player in repairing organs, NGF accelerates the process of growth, insulin-like growth factor serves importantly in the differentiation of stem cells into neuron-like cells. CONCLUSION ： As unipotent stem cells, NSCs have the abilities of self-renewal and potential of high differentiation. The method of mechanical dissociation is better than trypsin digestion in e separating ESCs. However, density gradient centrifuge separation is better than other methods in the separation of the BMSCs. NGF and other factors play an important role in the growth of NSCs.
基金supported by the Stem Cell Clinical Research Project of China,No.CMR-20161129-1003Liaoning Province Excellent Talent Program Project of China,No.XLYC1902031the Dalian Innovation Technology Foundation of China,No.2018J11CY025(all to JL).
文摘Biological studies typically rely on a simple monolayer cell culture,which does not reflect the complex functional characteristics of human tissues and organs,or their real response to external stimuli.Microfluidic technology has advantages of high-throughput screening,accurate control of the fluid velocity,low cell consumption,long-term culture,and high integration.By combining the multipotential differentiation of neural stem cells with high throughput and the integrated characteristics of microfluidic technology,an in vitro model of a functionalized neurovascular unit was established using human neural stem cell-derived neurons,astrocytes,oligodendrocytes,and a functional microvascular barrier.The model comprises a multi-layer vertical neural module and vascular module,both of which were connected with a syringe pump.This provides controllable conditions for cell inoculation and nutrient supply,and simultaneously simulates the process of ischemic/hypoxic injury and the process of inflammatory factors in the circulatory system passing through the blood-brain barrier and then acting on the nerve tissue in the brain.The in vitro functionalized neurovascular unit model will be conducive to central nervous system disease research,drug screening,and new drug development.
基金supported by grants of Jiangsu Natural College Foundation of China,No.13KJB310010,14KJB310015the Natural Foundation of Nantong University of China,No.14ZY022
文摘Fimbria-fornix transection induces both exogenous and endogenous neural stem cells to differentiate into neurons in the hippocampus.This indicates that the denervated hippocampus provides an environment for neuronal differentiation of neural stem cells.However,the pathways and mechanisms in this process are still unclear.Seven days after fimbria fornix transection,our reverse transcription polymerase chain reaction,western blot assay,and enzyme linked immunosorbent assay results show a significant increase in ciliary neurotrophic factor m RNA and protein expression in the denervated hippocampus.Moreover,neural stem cells derived from hippocampi of fetal（embryonic day 17） Sprague-Dawley rats were treated with ciliary neurotrophic factor for 7 days,with an increased number of microtubule associated protein-2-positive cells and decreased number of glial fibrillary acidic protein-positive cells detected.Our results show that ciliary neurotrophic factor expression is up-regulated in the denervated hippocampus,which may promote neuronal differentiation of neural stem cells in the denervated hippocampus.
基金support from Fundacao de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)+2 种基金the Provost’s Office for Research of the University of Sao Paulo, Grant number: 2011.1.9333.1.3 (NAPNA-USP)support from the German Research Council (IL 20/182, RI 2092/1-2, IL 20/21-1)the Sino-German Centre for the Support of Science (GZ 919)
文摘Purinergic receptors are among the first cell surface receptors expressed during embryonic development （Burnstock and Ulrich, 2011）. These are characterized based on their pharmacological properties of being activated by adenosine or purine/pyrimidine nucleotides as P1 and P2 receptors. P2 receptors are further classified by their structure as P2Y metabotropic and P2X ionotropic receptors.
基金supported by the National Natural Science Foundation of China,No.31340024
文摘Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.
基金supported by grants from the Spanish Ministerio de Cienciay Tecnología(BFU2007-67540)the Junta de Extremadura(PRI06A195,GR10152)
文摘Many blinding diseases,such as retinitis pigmentosa,age-related macular degeneration,and glaucoma involve the permanent loss of retinal neurons,especially photoreceptors or the centrally projecting retinal ganglion cells.Stem cells have been proposed as a potential source of cells for neuronal transplantation.
文摘Previous in vivo experiments have shown that human umbilical cord blood mesenchymal stem cells can promote the proliferation and differentiation of damaged celts, and help to repair damaged sites, Recent studies have reported that umbilical cord blood-derived mesenchymal stem cells can differentiate into neurons and glial cells. Recent studies have reported that the repair mechanisms underlying cord blood stern cells involve the replacement of damaged cells and mediation of the local micro-environment.
基金supported by the Army Laboratory Animal Foundation of China,No.SYDW[2020]22(to TC)the Shaanxi Provincial Key R&D Plan General Project of China,No.2022SF-236(to YM)the National Natural Science Foundation of China,No.82202070(to TC)。
文摘A microgravity environment has been shown to cause ocular damage and affect visual acuity,but the underlying mechanisms remain unclear.Therefore,we established an animal model of weightlessness via tail suspension to examine the pathological changes and molecular mechanisms of retinal damage under microgravity.After 4 weeks of tail suspension,there were no notable alterations in retinal function and morphology,while after 8 weeks of tail suspension,significant reductions in retinal function were observed,and the outer nuclear layer was thinner,with abundant apoptotic cells.To investigate the mechanism underlying the degenerative changes that occurred in the outer nuclear layer of the retina,proteomics was used to analyze differentially expressed proteins in rat retinas after 8 weeks of tail suspension.The results showed that the expression levels of fibroblast growth factor 2(also known as basic fibroblast growth factor)and glial fibrillary acidic protein,which are closely related to Müller cell activation,were significantly upregulated.In addition,Müller cell regeneration and Müller cell gliosis were observed after 4 and 8 weeks,respectively,of simulated weightlessness.These findings indicate that Müller cells play an important regulatory role in retinal outer nuclear layer degeneration during weightlessness.
基金supported by the National Key R&D Program of China(No.2018AAA0100300)the National Natural Science Foundation of China(Nos.82072018,82274375,and 82402490)+5 种基金the Anhui Provincial Science and Technology Major Project(No.202203a07020006)the Strategic Priority Research Program(C)of the Chinese Academy of Sciences(CAS)(No.XDC07040200)the Key R&D Program of Anhui Province(No.2022e07020012)the Natural Science Foundation of Anhui Province(No.2208085QH256)the Fundamental Research Funds for the Central Universities(No.WK2100000042)the China Postdoctoral Science Foundation(No.2022M713055).
文摘Neuroinflammation is the primary driver and signature of many neurodevelopmental disorders.However,because neurodevelopmental disorders caused by neuroinflammation are difficult to detect at the early stage,their progression remains unclear.To date,neither animal experiments nor in vitro models have uncovered their early developmental characteristics caused by neuroinflammation.In this study,we developed a neurovascular-unit-on-a-chip(NVU-on-a-chip)to model inflammation-induced neurodevelopmental disorders.With the chip,dynamic visualization of the progression caused by neuroinflammation was clearly demonstrated,and the changes in angiogenesis and neural differentiation under neuroinflammation were replicated.In addition,the activation of astrocytes and damage to neurons and capillaries at the early stage of neurodevelopmental disorders were observed.The results revealed for the first time the structural disruption of the neurovascular units and the neurovascular coupling failure caused by neuroinflammation.Furthermore,the outcomes of anti-inflammatory intervention using ibuprofen were preliminarily demonstrated.This work provides insights into the early progression of neurodevelopmental disorders caused by neuroinflammation and offers a platform for the development of therapeutic strategies for neuroinflammation.
基金supported by the National Key Research and Development Program of China,Nos.2017YFE0117000(to ZZQ),2018YFC1312302-3(to HQ)the Beijing Advanced Innovation Center for Intelligent Robots and Systems of China,No.2018IRS12(to ZZQ)the National Natural Science Foundation of China,Nos.82001167(to HL),81870844(to HQ),81701260(to ZZQ)。
文摘The mechanisms of the transplantation of neural stem cells(NSCs)in the treatment of Alzheimer’s disease remain poorly understood.In this study,NSCs were transplanted into the hippocampal CA1 region of the rTg(tau P301L)4510 mouse model,a tauopathy model that is thought to reflect the tau pathology associated with Alzheimer’s disease.The results revealed that NSC transplantation reduced the abnormal aggregation of tau,resulting in significant improvements in the short-term memory of the tauopathy model mice.Compared with wild-type and phosphate-buffered saline(PBS)-treated mice,mice that received NSC transplantations were characterized by changes in the expression of multiple proteins in brain tissue,particularly those related to the regulation of tau aggregation or misfolding.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis and Gene Ontology(GO)function analysis revealed that these proteins were primarily enriched in pathways associated with long-term potentiation,neurogenesis,and other neurobiological processes.Changes in the expression levels of key proteins were verified by western blot assays.These data provided clues to improve the understanding of the functional capacity associated with NSC transplantation in Alzheimer’s disease treatment.This study was approved by the Beijing Animal Ethics Association and Ethics Committee of Beijing Institute of Technology(approval No.SYXK-BIT-school of life science-2017-M03)in 2017.
基金This work was supported by National Stem Cell Clinical Research Registered Project,No.CMR-20161129-1003(to JL)Dalian Science and Technology Innovation Fund,No.2018J11CY025(to JL).
文摘Previous studies have shown that caveolin-1 is involved in regulating the differentiation of mesenchymal stem cells.However,its role in the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons remains unclear.The aim of this study was to investigate whether caveolin-1 regulates the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.We also examined whether the expression of caveolin-1 could be modulated by RNA interference technology to promote the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons.The differentiation of human adipose mesenchymal stem cells into dopaminergic neurons was evaluated morphologically and by examining expression of the markers tyrosine hydroxylase,Lmx1a and Nurr1.The analyses revealed that during the differentiation of human adipose mesenchymal stem cells into dopaminergic neurons,the expression of caveolin-1 is decreased.Notably,the downregulation of caveolin-1 promoted the differentiation of human adipose mesenchymal stem cells into dopaminergic-like neurons,and it increased the expression of tyrosine hydroxylase,Lmx1a and Nurr1.Together,our findings suggest that caveolin-1 plays a negative regulatory role in the differentiation of dopaminergic-like neurons from stem cells,and it may therefore be a potential molecular target for strategies for regulating the differentiation of these cells.This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Dalian Medical University of China(approval No.PJ-KS-KY-2020-54)on March 7,2017.
基金sponsored by Shanghai Key Projects of Basic Research,No.08JC1413900
文摘The difference between Noggin and basic fibroblast growth factor for the neural precursor differen- tiation from human embryonic stem cells has not been studied. In this study, 100 tJg/L Noggin or 20 IJg/L basic fibroblast growth factor in serum-free neural induction medium was used to differen- tiate human embryonic stem cells H14 into neural precursors using monolayer differentiation. Two weeks after induction, significantly higher numbers of neural rosettes formed in the Noggin-induced group than the basic fibroblast growth factor-induced group, as detected by phase contrast micro- scope. Immunofluorescence staining revealed expression levels of Nestin, [3-111 Tubulin and Sox-1 were higher in the induced cells and reverse-transcription PCR showed induced cells expressed Nestin, Sox-1 and Neurofilament mRNA. Protein and mRNA expression in the Noggin-induced group was increased compared with the basic fibroblast growth factor-induced group. Noggin has a greater effect than basic fibroblast growth factor on the induction of human embryonic stem cell differentiation into neural precursors by monolayer differentiation, as Noggin accelerates and in- creases the differentiation of neural precursors.
