Skeletal muscle-derived cells have strong secretory function,while skeletal muscle-derived stem cells,which are included in muscle-derived cells,can differentiate into Schwann cell-like cells and other cell types.Howe...Skeletal muscle-derived cells have strong secretory function,while skeletal muscle-derived stem cells,which are included in muscle-derived cells,can differentiate into Schwann cell-like cells and other cell types.However,the effect of muscle-derived cells on peripheral nerve defects has not been reported.In this study,5-mm-long nerve defects were created in the right sciatic nerves of mice to construct a peripheral nerve defect model.Adult female C57BL/6 mice were randomly divided into four groups.For the muscle-derived cell group,muscle-derived cells were injected into the catheter after the cut nerve ends were bridged with a polyurethane catheter.For external oblique muscle-fabricated nerve conduit and polyurethane groups,an external oblique muscle-fabricated nerve conduit or polyurethane catheter was used to bridge the cut nerve ends,respectively.For the sham group,the sciatic nerves on the right side were separated but not excised.At 8 and 12 weeks post-surgery,distributions of axons and myelin sheaths were observed,and the nerve diameter was calculated using immunofluorescence staining.The number,diameter,and thickness of myelinated nerve fibers were detected by toluidine blue staining and transmission electron microscopy.Muscle fiber area ratios were calculated by Masson’s trichrome staining of gastrocnemius muscle sections.Sciatic functional index was recorded using walking footprint analysis at 4,8,and 12 weeks after operation.The results showed that,at 8 and 12 weeks after surgery,myelin sheaths and axons of regenerating nerves were evenly distributed in the muscle-derived cell group.The number,diameter,and myelin sheath thickness of myelinated nerve fibers,as well as gastrocnemius muscle wet weight and muscle area ratio,were significantly higher in the muscle-derived cell group compared with the polyurethane group.At 4,8,and 12 weeks post-surgery,sciatic functional index was notably increased in the muscle-derived cell group compared with the polyurethane group.These criteria of the muscle-derived cell group were not significantly different from the external oblique muscle-fabricated nerve conduit group.Collectively,these data suggest that muscle-derived cells effectively accelerated peripheral nerve regeneration.This study was approved by the Animal Ethics Committee of Plastic Surgery Hospital,Chinese Academy of Medical Sciences(approval No.040)on September 28,2016.展开更多
An overview of a long-gap peripheral nerve therapy： A long- gap peripheral nerve transection injury is an irreparable injury to the living body, and mostly leads to permanent loss of re- lated motor and sensory funct...An overview of a long-gap peripheral nerve therapy： A long- gap peripheral nerve transection injury is an irreparable injury to the living body, and mostly leads to permanent loss of re- lated motor and sensory functions. In such long gap injuries, nerve end-to-end suture is physically impossible. Therefore, bridging a long nerve-gap is critical to re-establish adequate mechanical support for separated nerve ends, and prevent the diffusion of neurotrophic and neurotropic factors secreted by transected stumps （Deumens et al., 2010）.展开更多
Muscle-derived stem cells were isolated from the skeletal muscle of Sprague-Dawley neonatal rats aged 3 days old. Cells at passage 5 were incubated in Dulbecco＇s modified Eagle＇s medium supplemented with 10% （v/v）...Muscle-derived stem cells were isolated from the skeletal muscle of Sprague-Dawley neonatal rats aged 3 days old. Cells at passage 5 were incubated in Dulbecco＇s modified Eagle＇s medium supplemented with 10% （v/v） fetal bovine serum, 20 IJg/L nerve growth factor, 20 pg/L basic fibroblast growth factor and 1% （v/v） penicillin for 6 days. Cells presented with long processes, similar to nerve cells. Connections were formed between cell processes. Immunocytochemical staining with neuron specific enolase verified that cells differentiated into neuron-like cells. Immunofluorescence cytochemistry and western blot results revealed that the expression of protein hairy enhancer of split-1 was significantly reduced. These results indicate that low expression of protein hairy enhancer of split-1 participates in the differentiation of muscle-derived stem cells into neuron-like cells.展开更多
OBJECTIVE： To identify global research trends of muscle-derived stem cells （MDSCs） using a bibliometric analysis of the Web of Science, Research Portfolio Online Reporting Tools of the National Institutes of Health...OBJECTIVE： To identify global research trends of muscle-derived stem cells （MDSCs） using a bibliometric analysis of the Web of Science, Research Portfolio Online Reporting Tools of the National Institutes of Health （NIH）, and the Clinical Trials registry database （ClinicalTrials.gov）. DATA RETRIEVAL： We performed a bibliometric analysis of data retrievals for MDSCs from 2002 to 2011 using the Web of Science, NIH, and ClinicalTrials.gov. SELECTION CRITERIA： Inclusion criteria： （1） Web of Science： （a） peer-reviewed articles on MDSCs that were published and indexed in the Web of Science. （b） Type of articles： original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material and news items. （c） Year of publication： 2002-2011. （d） Citation databases： Science Citation Index-Expanded （SCI-E）, 1899-present; Conference Proceedings Citation Index-Science （CPCI-S）, 1991-present; Book Citation Index-Science （BKCI-S）, 2005-present. （2） NIH： （a） Projects on MDSCs supported by the NIH. （b） Fiscal year： 1988-present. （3） ClinicalTrials.gov： All clinical trials relating to MDSCs were searched in this database. Exclusion criteria： （1） Web of Science： （a） Articles that required manual searching or telephone access. （b） We excluded documents that were not published in the public domain. （c） We excluded a number of corrected papers from the total number of articles. （d） We excluded articles from the following databases： Social Sciences Citation Index （SSCI）, 1898-present; Arts ＆ Humanities Citation Index （A＆HCI）, 1975-present; Conference Proceedings Citation Index - Social Science ＆ Humanities （CPCI-SSH）, 1991-present; Book Citation Index - Social Sciences ＆ Humanities （BKCI-SSH）, 2005-present; Current Chemical Reactions （CCR-EXPANDED）, 1985-present; Index Chemicus （IC）, 1993-present. （2） NIH： （a） We excluded publications related to MDSCs that were supported by the NIH. （b） We limited the keyword search to studies that included MDSCs within the title or abstract. （3） ClinicalTrials.gov： （a） We excluded clinical trials that were not in the ClinicalTrials.gov database. （b） We excluded clinical trials that dealt with stem cells other than MDSCs in the ClinicalTrials.gov database. MAIN OUTCOME MEASURES： （1） Type of literature; （2） annual publication output; （3） distribution according to journals; （4） distribution according to country; （5） distribution according to institution; （6） top cited authors over the last 10 years; （7） projects financially supported by the NIH; and （8） clinical trials registered. RESULTS： （1） In all, 802 studies on MDSCs appeared in the Web of Science from 2002 to 2011, almost half of which derived from American authors and institutes. The number of studies on MDSCs has gradually increased over the past 10 years. Most papers on MDSCs appeared in journals with a particular focus on cell biology research, such as Experimental Cell Research, Journal of Cell Science, and PLoS One. （2） Eight MDSC research projects have received over US$6 billion in funding from the NIH. The current project led by Dr. Johnny Huard of the University of Pittsburgh-＂Muscle-Based Tissue Engineering to Improve Bone Healing＂-is supported by the NIH. Dr. Huard has been the most productive and top-cited author in the field of gene therapy and adult stem cell research in the Web of Science over last 10 years. （3） On ClinicalTrials.gov, ＂Muscle Derived Cell Therapy for Bladder Exstrophy Epispadias Induced Incontinence＂ Phase 1 is registered and sponsored by Johns Hopkins University and has been led by Dr. John P. Gearhart since November 2009. CONCLUSION： From our analysis of the literature and research trends, we found that MDSCs may offer further benefits in regenerative medicine.展开更多
We cultured rat muscle-derived stem cells in medium containing nerve growth factor and basic fi-broblast growth factor to induce neuronal-like cell differentiation.Immunocytochemical staining and reverse transcription...We cultured rat muscle-derived stem cells in medium containing nerve growth factor and basic fi-broblast growth factor to induce neuronal-like cell differentiation.Immunocytochemical staining and reverse transcription-PCR showed that the differentiated muscle-derived stem cells exhibited processes similar to those of neuronal-like cells and neuron-specific enolase expression,but Notch1 mRNA and protein expression was decreased.Down-regulation of Notch1 expression may facilitate neuronal-like cell differentiation from muscle-derived stem cells.展开更多
Muscle-derived stem cells （MDSCs） can come from a number of different sources, which are easy to isolate and culture, and are also useful in the transformation and expression of exogenous genes. Therefore, MDSCs cou...Muscle-derived stem cells （MDSCs） can come from a number of different sources, which are easy to isolate and culture, and are also useful in the transformation and expression of exogenous genes. Therefore, MDSCs could possibly be used for gene therapy in the treatment of neurological diseases. However, research on MDSCs has focused on identifying phenotypes and induced differentiation, with few in vivo animal experiments conducted. In this study, MDSCs were selected as seed cells and implanted into the rat spinal cord injury area. Results demonstrated that the MDSCs survived, migrated, and were distributed along the spinal nerves. Moreover, the motor function of rat lower limbs improved significantly, suggesting that MDSCs could be used as seed cells to repair spinal cord injury.展开更多
Adult stem cells from skeletal muscle cells were induced to differentiate into cardiocytes to see if stem cells from another different but histologically-comparable tissues can differentiate to the target cells. Skele...Adult stem cells from skeletal muscle cells were induced to differentiate into cardiocytes to see if stem cells from another different but histologically-comparable tissues can differentiate to the target cells. Skeletal muscles-derived stem cells （MDSCs） were isolated from adult skeleton muscle tissues by differential adhesion, and immunocytochemically identified by using Sca-1. In order to induce the proliferation but not differentiation of MDSCs, the cells were cultured in Dulbecco’s modified Eagle’s medium/F12 （DMEM/F12） supplemented with 1：50 B27, 20 ng/mL basic fibroblast growth factor （bFGF）, 20 ng/mL epidermal growth factor （EGF） in a suspension for 6 days. Then these stem cells were treated with 5 μmol/L 5-azacytidine for 24 h in an adherence culture. The characteristics of induced cells were examined by immunocytochemistry, quantitative real time RT-PCR and morphological observation of cell phenotype. Our results showed that the appearance of some cells gradually changed from spindle-shape into polygonal or short-column-shape. Some of these post-treated cells could contract spontaneously and rhythmically. The expression of GATA-4 and cTnT was increased 1 and 2 week（s） after the treatment. And about 16.6% of post-treated cells were cTnT-positive. Therefore, we are led to conclude that skeletal muscle-derived stem cells could differentiate into cardiocyte-like cells, which exhibited some characteristics of cardiocytes.展开更多
The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells （MDSCs）. Rat skeletal MDSCs were obtained by using preplate techn...The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells （MDSCs）. Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups： group A （control group）, group B （treated with TGF-β1, 10 ng/rnL）, group C （treated with TGF-β1 and anti-connective tissue growth factor （CTGF）, both in 10 ng/mL）, and group D （treated with anti-CTGF, 10 ng/mL）. The expression of CTGF, collagen type- I （COL- I ） and collagen type-III （COL-III） in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL- I and COL-Ⅲ was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL- I and COL-Ⅲ mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-Ill induced by TGF-131 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL- I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL- I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.展开更多
Background Regular exercise training provides significant health benefits among cancer survivors and is associated with lower breast cancer mortality and reduced risk of recurrence.Both exercise-induced factors secret...Background Regular exercise training provides significant health benefits among cancer survivors and is associated with lower breast cancer mortality and reduced risk of recurrence.Both exercise-induced factors secreted into circulation(exerkines)and bioactive molecules contained in skeletal muscle secretome have been proposed to affect the tumor microenvironment and mediate some of the anti-carcinogenic effects of exercise.This study utilized exercise-conditioned human serum obtained from breast cancer patients during chemotherapy and skeletal myotubes’secretome after mechanical loading to investigate their effects on breast cancer cells in vitro.Methods Breast cancer patients participated in a 12-week exercise training program during their chemotherapy,and blood serum was collected immediately before and after an exercise session in the 2nd and 12th weeks of training.Skeletal myoblasts were differentiated into myotubes and subjected to mechanical stretching to collect their secretome(stretch medium(SM)).Hormone-sensitive Michigan Cancer Foundation-7(MCF-7)and triple-negative M.D.Anderson-Metastatic Breast-231(MDA-MB-231)breast cancer cells were treated with either human serum or with the skeletal myotubes’secretome to examine their metabolic activity,migration,cytotoxicity levels and apoptosis regulation.Results The exercise-conditioned serum obtained from breast cancer patients who were subjected to the 12-week training during chemotherapy resulted in reduced metabolic activity(p<0.001)and increased lactate dehydrogenase activity(cytotoxicity)(p<0.001)in both MCF-7 and MDA-MB-231 breast cancer cells when compared with the control condition.Moreover,incubation of breast cancer cells with the post-exercise serum induced apoptosis in MCF-7 and MDA-MB-231 cells,as indicated by increase in DNA damage and the percentage of necrotic cells(p<0.05)when compared to pre-exercise condition.Similarly,a significant decrease(p<0.001)was observed in the metabolic activity of MCF-7 cells treated with the SM,along with increased cytotoxicity(p<0.05),compared to the cells cultured with the regular growth media.Comparable though not as profound effects were observed in MDA-MB-231 cells when treated with the SM secretome.Furthermore,the expression of apoptosis-inducing Caspase-7(p<0.001)and Caspase-8(p<0.01)proteins was increased,whereas cell survival-regulating factors interleukin-8(IL-8)(p<0.001),superoxide dismutase-2(p<0.05),Fas cell surface death receptor(p<0.05),and vascular endothelial growth factor(p<0.01)were downregulated in the SM-treated MCF-7 cells.In addition,the migrating behavior of MCF-7 cells was diminished,and higher levels of DNA damage were observed in cells treated with either SM or non-stretch media.Conclusion Both exercise-conditioned serum of breast cancer patients and skeletal myotubes secretome after mechanical loading can reduce the metabolic activity,promote cell toxicity and DNA damage,modulate the protein expression of crucial cell survival-regulating factors,and lead to apoptosis in breast cancer cells.These findings suggest that even after cancer diagnosis,exercise may exert beneficial effects additive to chemotherapy against breast cancer prognosis.展开更多
BACKGROUND Mesenchymal stromal/stem cells (MSCs) constitute a promising tool in regenerative medicine and can be isolated from different human tissues. However, their biological properties are still not fully characte...BACKGROUND Mesenchymal stromal/stem cells (MSCs) constitute a promising tool in regenerative medicine and can be isolated from different human tissues. However, their biological properties are still not fully characterized. Whereas MSCs from different tissue exhibit many common characteristics, their biological activity and some markers are different and depend on their tissue of origin. Understanding the factors that underlie MSC biology should constitute important points for consideration for researchers interested in clinical MSC application. AIM To characterize the biological activity of MSCs during longterm culture isolated from: bone marrow (BM-MSCs), adipose tissue (AT-MSCs), skeletal muscles (SMMSCs), and skin (SK-MSCs). METHODS MSCs were isolated from the tissues, cultured for 10 passages, and assessed for: phenotype with immunofluorescence and flow cytometry, multipotency with differentiation capacity for osteo-, chondro-, and adipogenesis, stemness markers with qPCR for mRNA for Sox2 and Oct4, and genetic stability for p53 and c-Myc;27 bioactive factors were screened using the multiplex ELISA array, and spontaneous fusion involving a co-culture of SM-MSCs with BM-MSCs or AT-MSCs stained with PKH26 (red) or PKH67 (green) was performed. RESULTS All MSCs showed the basic MSC phenotype;however, their expression decreased during the follow-up period, as confirmed by fluorescence intensity. The examined MSCs express CD146 marker associated with proangiogenic properties;however their expression decreased in AT-MSCs and SM-MSCs, but was maintained in BM-MSCs. In contrast, in SK-MSCs CD146 expression increased in late passages. All MSCs, except BM-MSCs, expressed PW1, a marker associated with differentiation capacity and apoptosis. BM-MSCs and AT-MSCs expressed stemness markers Sox2 and Oct4 in long-term culture. All MSCs showed a stable p53 and c-Myc expression. BM-MSCs and AT-MSCs maintained their differentiation capacity during the follow-up period. In contrast, SK-MSCs and SM-MSCs had a limited ability to differentiate into adipocytes. BM-MSCs and AT-MSCs revealed similarities in phenotype maintenance, capacity for multilineage differentiation, and secretion of bioactive factors. Because AT-MSCs fused with SM-MSCs as effectively as BM-MSCs, AT-MSCs may constitute an alternative source for BM-MSCs. CONCLUSION Long-term culture affects the biological activity of MSCs obtained from various tissues. The source of MSCs and number of passages are important considerations in regenerative medicine.展开更多
Stress urinary incontinence is one of the most common diseases in urology. The main treatments for stress urinary incontinence are pharmacotherapy, physicobehavioral therapy and surgery.However, the results of present...Stress urinary incontinence is one of the most common diseases in urology. The main treatments for stress urinary incontinence are pharmacotherapy, physicobehavioral therapy and surgery.However, the results of present methods are not satisfactory. Tissue engineering is a newly emerging technology that may provide a novel method for the treatment of stress urinary incontinence.展开更多
基金financially supported by the National Natural Science Foundation of China,No.81671908(to ZLQ)and No.81571921(to XNY)the Fundamental Research Fund for the Central Universities of China,No.2016ZX310197(to ZLQ)+1 种基金the Union Youth Science&Research Foundation of China,No.3332015155(to XNY)the Science Fund of Plastic Surgery Hospital,Chinese Academy of Medical Sciences,and Peking Union Medical College of China,No.Q2015013(to XNY)
文摘Skeletal muscle-derived cells have strong secretory function,while skeletal muscle-derived stem cells,which are included in muscle-derived cells,can differentiate into Schwann cell-like cells and other cell types.However,the effect of muscle-derived cells on peripheral nerve defects has not been reported.In this study,5-mm-long nerve defects were created in the right sciatic nerves of mice to construct a peripheral nerve defect model.Adult female C57BL/6 mice were randomly divided into four groups.For the muscle-derived cell group,muscle-derived cells were injected into the catheter after the cut nerve ends were bridged with a polyurethane catheter.For external oblique muscle-fabricated nerve conduit and polyurethane groups,an external oblique muscle-fabricated nerve conduit or polyurethane catheter was used to bridge the cut nerve ends,respectively.For the sham group,the sciatic nerves on the right side were separated but not excised.At 8 and 12 weeks post-surgery,distributions of axons and myelin sheaths were observed,and the nerve diameter was calculated using immunofluorescence staining.The number,diameter,and thickness of myelinated nerve fibers were detected by toluidine blue staining and transmission electron microscopy.Muscle fiber area ratios were calculated by Masson’s trichrome staining of gastrocnemius muscle sections.Sciatic functional index was recorded using walking footprint analysis at 4,8,and 12 weeks after operation.The results showed that,at 8 and 12 weeks after surgery,myelin sheaths and axons of regenerating nerves were evenly distributed in the muscle-derived cell group.The number,diameter,and myelin sheath thickness of myelinated nerve fibers,as well as gastrocnemius muscle wet weight and muscle area ratio,were significantly higher in the muscle-derived cell group compared with the polyurethane group.At 4,8,and 12 weeks post-surgery,sciatic functional index was notably increased in the muscle-derived cell group compared with the polyurethane group.These criteria of the muscle-derived cell group were not significantly different from the external oblique muscle-fabricated nerve conduit group.Collectively,these data suggest that muscle-derived cells effectively accelerated peripheral nerve regeneration.This study was approved by the Animal Ethics Committee of Plastic Surgery Hospital,Chinese Academy of Medical Sciences(approval No.040)on September 28,2016.
文摘An overview of a long-gap peripheral nerve therapy： A long- gap peripheral nerve transection injury is an irreparable injury to the living body, and mostly leads to permanent loss of re- lated motor and sensory functions. In such long gap injuries, nerve end-to-end suture is physically impossible. Therefore, bridging a long nerve-gap is critical to re-establish adequate mechanical support for separated nerve ends, and prevent the diffusion of neurotrophic and neurotropic factors secreted by transected stumps （Deumens et al., 2010）.
基金supported by a grant from the Distinguished Young Scholars in Universities in Liaoning Province Development Plan, No. LJQ2011090
文摘Muscle-derived stem cells were isolated from the skeletal muscle of Sprague-Dawley neonatal rats aged 3 days old. Cells at passage 5 were incubated in Dulbecco＇s modified Eagle＇s medium supplemented with 10% （v/v） fetal bovine serum, 20 IJg/L nerve growth factor, 20 pg/L basic fibroblast growth factor and 1% （v/v） penicillin for 6 days. Cells presented with long processes, similar to nerve cells. Connections were formed between cell processes. Immunocytochemical staining with neuron specific enolase verified that cells differentiated into neuron-like cells. Immunofluorescence cytochemistry and western blot results revealed that the expression of protein hairy enhancer of split-1 was significantly reduced. These results indicate that low expression of protein hairy enhancer of split-1 participates in the differentiation of muscle-derived stem cells into neuron-like cells.
基金supported by the National Natural Science Foundation of China, No. 30772190Creative Group Project from Education Department of Liaoning Province, No.2009T063
文摘OBJECTIVE： To identify global research trends of muscle-derived stem cells （MDSCs） using a bibliometric analysis of the Web of Science, Research Portfolio Online Reporting Tools of the National Institutes of Health （NIH）, and the Clinical Trials registry database （ClinicalTrials.gov）. DATA RETRIEVAL： We performed a bibliometric analysis of data retrievals for MDSCs from 2002 to 2011 using the Web of Science, NIH, and ClinicalTrials.gov. SELECTION CRITERIA： Inclusion criteria： （1） Web of Science： （a） peer-reviewed articles on MDSCs that were published and indexed in the Web of Science. （b） Type of articles： original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material and news items. （c） Year of publication： 2002-2011. （d） Citation databases： Science Citation Index-Expanded （SCI-E）, 1899-present; Conference Proceedings Citation Index-Science （CPCI-S）, 1991-present; Book Citation Index-Science （BKCI-S）, 2005-present. （2） NIH： （a） Projects on MDSCs supported by the NIH. （b） Fiscal year： 1988-present. （3） ClinicalTrials.gov： All clinical trials relating to MDSCs were searched in this database. Exclusion criteria： （1） Web of Science： （a） Articles that required manual searching or telephone access. （b） We excluded documents that were not published in the public domain. （c） We excluded a number of corrected papers from the total number of articles. （d） We excluded articles from the following databases： Social Sciences Citation Index （SSCI）, 1898-present; Arts ＆ Humanities Citation Index （A＆HCI）, 1975-present; Conference Proceedings Citation Index - Social Science ＆ Humanities （CPCI-SSH）, 1991-present; Book Citation Index - Social Sciences ＆ Humanities （BKCI-SSH）, 2005-present; Current Chemical Reactions （CCR-EXPANDED）, 1985-present; Index Chemicus （IC）, 1993-present. （2） NIH： （a） We excluded publications related to MDSCs that were supported by the NIH. （b） We limited the keyword search to studies that included MDSCs within the title or abstract. （3） ClinicalTrials.gov： （a） We excluded clinical trials that were not in the ClinicalTrials.gov database. （b） We excluded clinical trials that dealt with stem cells other than MDSCs in the ClinicalTrials.gov database. MAIN OUTCOME MEASURES： （1） Type of literature; （2） annual publication output; （3） distribution according to journals; （4） distribution according to country; （5） distribution according to institution; （6） top cited authors over the last 10 years; （7） projects financially supported by the NIH; and （8） clinical trials registered. RESULTS： （1） In all, 802 studies on MDSCs appeared in the Web of Science from 2002 to 2011, almost half of which derived from American authors and institutes. The number of studies on MDSCs has gradually increased over the past 10 years. Most papers on MDSCs appeared in journals with a particular focus on cell biology research, such as Experimental Cell Research, Journal of Cell Science, and PLoS One. （2） Eight MDSC research projects have received over US$6 billion in funding from the NIH. The current project led by Dr. Johnny Huard of the University of Pittsburgh-＂Muscle-Based Tissue Engineering to Improve Bone Healing＂-is supported by the NIH. Dr. Huard has been the most productive and top-cited author in the field of gene therapy and adult stem cell research in the Web of Science over last 10 years. （3） On ClinicalTrials.gov, ＂Muscle Derived Cell Therapy for Bladder Exstrophy Epispadias Induced Incontinence＂ Phase 1 is registered and sponsored by Johns Hopkins University and has been led by Dr. John P. Gearhart since November 2009. CONCLUSION： From our analysis of the literature and research trends, we found that MDSCs may offer further benefits in regenerative medicine.
基金Program for Liaoning Innovative Research Team in University(LNIRT),No.2008T113
文摘We cultured rat muscle-derived stem cells in medium containing nerve growth factor and basic fi-broblast growth factor to induce neuronal-like cell differentiation.Immunocytochemical staining and reverse transcription-PCR showed that the differentiated muscle-derived stem cells exhibited processes similar to those of neuronal-like cells and neuron-specific enolase expression,but Notch1 mRNA and protein expression was decreased.Down-regulation of Notch1 expression may facilitate neuronal-like cell differentiation from muscle-derived stem cells.
基金the Natural Science Foundation of Liaoning Province,No.20072204
文摘Muscle-derived stem cells （MDSCs） can come from a number of different sources, which are easy to isolate and culture, and are also useful in the transformation and expression of exogenous genes. Therefore, MDSCs could possibly be used for gene therapy in the treatment of neurological diseases. However, research on MDSCs has focused on identifying phenotypes and induced differentiation, with few in vivo animal experiments conducted. In this study, MDSCs were selected as seed cells and implanted into the rat spinal cord injury area. Results demonstrated that the MDSCs survived, migrated, and were distributed along the spinal nerves. Moreover, the motor function of rat lower limbs improved significantly, suggesting that MDSCs could be used as seed cells to repair spinal cord injury.
基金supported by a grant from the National Natural Sciences Foundation of China (No 30872627)Hubei Provincial Natural Sciences Foundation (No 2007ABA133)
文摘Adult stem cells from skeletal muscle cells were induced to differentiate into cardiocytes to see if stem cells from another different but histologically-comparable tissues can differentiate to the target cells. Skeletal muscles-derived stem cells （MDSCs） were isolated from adult skeleton muscle tissues by differential adhesion, and immunocytochemically identified by using Sca-1. In order to induce the proliferation but not differentiation of MDSCs, the cells were cultured in Dulbecco’s modified Eagle’s medium/F12 （DMEM/F12） supplemented with 1：50 B27, 20 ng/mL basic fibroblast growth factor （bFGF）, 20 ng/mL epidermal growth factor （EGF） in a suspension for 6 days. Then these stem cells were treated with 5 μmol/L 5-azacytidine for 24 h in an adherence culture. The characteristics of induced cells were examined by immunocytochemistry, quantitative real time RT-PCR and morphological observation of cell phenotype. Our results showed that the appearance of some cells gradually changed from spindle-shape into polygonal or short-column-shape. Some of these post-treated cells could contract spontaneously and rhythmically. The expression of GATA-4 and cTnT was increased 1 and 2 week（s） after the treatment. And about 16.6% of post-treated cells were cTnT-positive. Therefore, we are led to conclude that skeletal muscle-derived stem cells could differentiate into cardiocyte-like cells, which exhibited some characteristics of cardiocytes.
基金supported by the National Natural Science Foundation of China(No.30872627)
文摘The aim of this study was to investigate the mechanism of deposition of extracellular matrix induced by TGF-β1 in skeletal muscle-derived stem cells （MDSCs）. Rat skeletal MDSCs were obtained by using preplate technique, and divided into four groups： group A （control group）, group B （treated with TGF-β1, 10 ng/rnL）, group C （treated with TGF-β1 and anti-connective tissue growth factor （CTGF）, both in 10 ng/mL）, and group D （treated with anti-CTGF, 10 ng/mL）. The expression of CTGF, collagen type- I （COL- I ） and collagen type-III （COL-III） in MDSCs was examined by using RT-PCR, Western blot and immunofluorescent stain. It was found that one day after TGF-β1 treatment, the expression of CTGF, COL- I and COL-Ⅲ was increased dramatically. CTGF expression reached the peak on the day 2, and then decreased rapidly to a level of control group on the day 5. COL- I and COL-Ⅲ mRNA levels were overexpresed on the day 2 and 3 respectively, while their protein expression levels were up-regulated on the day 2 and reached the peak on the day 7. In group C, anti-CTGF could partly suppress the overexpression of COL-I and COL-Ill induced by TGF-131 one day after adding CTGF antibody. It was concluded that TGF-β1 could induce MDSCs to express CTGF, and promote the production of COL- I and COL-III. In contrast, CTGF antibody could partially inhibit the effect of TGF-β1 on the MDSCs by reducing the expression of COL- I and COL-III. Taken together, we demonstrated that TGF-β1-CTGF signaling played a crucial role in MDSCs synthesizing collagen proteins in vitro, which provided theoretical basis for exploring the methods postponing skeletal muscle fibrosis after nerve injury.
基金the Biochemical Imaging Center at UmeåUniversity and the National Microscopy Infrastructure(VR-RFI 2019-00217),for providing assistance in microscopy.
文摘Background Regular exercise training provides significant health benefits among cancer survivors and is associated with lower breast cancer mortality and reduced risk of recurrence.Both exercise-induced factors secreted into circulation(exerkines)and bioactive molecules contained in skeletal muscle secretome have been proposed to affect the tumor microenvironment and mediate some of the anti-carcinogenic effects of exercise.This study utilized exercise-conditioned human serum obtained from breast cancer patients during chemotherapy and skeletal myotubes’secretome after mechanical loading to investigate their effects on breast cancer cells in vitro.Methods Breast cancer patients participated in a 12-week exercise training program during their chemotherapy,and blood serum was collected immediately before and after an exercise session in the 2nd and 12th weeks of training.Skeletal myoblasts were differentiated into myotubes and subjected to mechanical stretching to collect their secretome(stretch medium(SM)).Hormone-sensitive Michigan Cancer Foundation-7(MCF-7)and triple-negative M.D.Anderson-Metastatic Breast-231(MDA-MB-231)breast cancer cells were treated with either human serum or with the skeletal myotubes’secretome to examine their metabolic activity,migration,cytotoxicity levels and apoptosis regulation.Results The exercise-conditioned serum obtained from breast cancer patients who were subjected to the 12-week training during chemotherapy resulted in reduced metabolic activity(p<0.001)and increased lactate dehydrogenase activity(cytotoxicity)(p<0.001)in both MCF-7 and MDA-MB-231 breast cancer cells when compared with the control condition.Moreover,incubation of breast cancer cells with the post-exercise serum induced apoptosis in MCF-7 and MDA-MB-231 cells,as indicated by increase in DNA damage and the percentage of necrotic cells(p<0.05)when compared to pre-exercise condition.Similarly,a significant decrease(p<0.001)was observed in the metabolic activity of MCF-7 cells treated with the SM,along with increased cytotoxicity(p<0.05),compared to the cells cultured with the regular growth media.Comparable though not as profound effects were observed in MDA-MB-231 cells when treated with the SM secretome.Furthermore,the expression of apoptosis-inducing Caspase-7(p<0.001)and Caspase-8(p<0.01)proteins was increased,whereas cell survival-regulating factors interleukin-8(IL-8)(p<0.001),superoxide dismutase-2(p<0.05),Fas cell surface death receptor(p<0.05),and vascular endothelial growth factor(p<0.01)were downregulated in the SM-treated MCF-7 cells.In addition,the migrating behavior of MCF-7 cells was diminished,and higher levels of DNA damage were observed in cells treated with either SM or non-stretch media.Conclusion Both exercise-conditioned serum of breast cancer patients and skeletal myotubes secretome after mechanical loading can reduce the metabolic activity,promote cell toxicity and DNA damage,modulate the protein expression of crucial cell survival-regulating factors,and lead to apoptosis in breast cancer cells.These findings suggest that even after cancer diagnosis,exercise may exert beneficial effects additive to chemotherapy against breast cancer prognosis.
基金the National Science Center,No.N407121940the Wroclaw Centre of Biotechnology,the Leading National Research Centre(KNOW)program for the years 2014-2018
文摘BACKGROUND Mesenchymal stromal/stem cells (MSCs) constitute a promising tool in regenerative medicine and can be isolated from different human tissues. However, their biological properties are still not fully characterized. Whereas MSCs from different tissue exhibit many common characteristics, their biological activity and some markers are different and depend on their tissue of origin. Understanding the factors that underlie MSC biology should constitute important points for consideration for researchers interested in clinical MSC application. AIM To characterize the biological activity of MSCs during longterm culture isolated from: bone marrow (BM-MSCs), adipose tissue (AT-MSCs), skeletal muscles (SMMSCs), and skin (SK-MSCs). METHODS MSCs were isolated from the tissues, cultured for 10 passages, and assessed for: phenotype with immunofluorescence and flow cytometry, multipotency with differentiation capacity for osteo-, chondro-, and adipogenesis, stemness markers with qPCR for mRNA for Sox2 and Oct4, and genetic stability for p53 and c-Myc;27 bioactive factors were screened using the multiplex ELISA array, and spontaneous fusion involving a co-culture of SM-MSCs with BM-MSCs or AT-MSCs stained with PKH26 (red) or PKH67 (green) was performed. RESULTS All MSCs showed the basic MSC phenotype;however, their expression decreased during the follow-up period, as confirmed by fluorescence intensity. The examined MSCs express CD146 marker associated with proangiogenic properties;however their expression decreased in AT-MSCs and SM-MSCs, but was maintained in BM-MSCs. In contrast, in SK-MSCs CD146 expression increased in late passages. All MSCs, except BM-MSCs, expressed PW1, a marker associated with differentiation capacity and apoptosis. BM-MSCs and AT-MSCs expressed stemness markers Sox2 and Oct4 in long-term culture. All MSCs showed a stable p53 and c-Myc expression. BM-MSCs and AT-MSCs maintained their differentiation capacity during the follow-up period. In contrast, SK-MSCs and SM-MSCs had a limited ability to differentiate into adipocytes. BM-MSCs and AT-MSCs revealed similarities in phenotype maintenance, capacity for multilineage differentiation, and secretion of bioactive factors. Because AT-MSCs fused with SM-MSCs as effectively as BM-MSCs, AT-MSCs may constitute an alternative source for BM-MSCs. CONCLUSION Long-term culture affects the biological activity of MSCs obtained from various tissues. The source of MSCs and number of passages are important considerations in regenerative medicine.
文摘Stress urinary incontinence is one of the most common diseases in urology. The main treatments for stress urinary incontinence are pharmacotherapy, physicobehavioral therapy and surgery.However, the results of present methods are not satisfactory. Tissue engineering is a newly emerging technology that may provide a novel method for the treatment of stress urinary incontinence.
