BACKGROUND： Midbrain-derived neural stem cells （mNSCs） can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson＇s disease. OBJECTIVE： ...BACKGROUND： Midbrain-derived neural stem cells （mNSCs） can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson＇s disease. OBJECTIVE： To isolate rat embryonic mNSCs and to observe the differentiation characteristics of mNSCs induced by cell growth-promoting factors. DESIGN, TIME AND SETTING： An in vitro cell culture study based on the molecular biology of nerve cells was carried out at the Institute of Clinical Medicine, China-Japan Friendship Hospital （China） from March to November 2007. MATERIALS： Sprague Dawley rats at embryonic day 14 were used in this study. Nestin antibody, β-Ⅲ tubulin antibody, glial fibrillary acidic protein （GFAP） antibody and cyclic nucleotide 3＇-phosphohydrolase （CNPase） antibody were provided by Abcam; DMEM/F12 medium and N2 supplement were provided by Invitrogen; epidermal growth factor （EGF） and fibroblast growth factor-2 （FGF2） were provided by R＆D Systems. METHODS： The ventral mesencephalon was dissected from embryonic day 14 rat embryos. By trypsin digestion and mechanical separation, the brain tissue was triturated into a fine single-cell suspension. The cells were cultured in 5 mL serum-free medium containing DMEM/FI 2, 1% N： supplement, 20 ng/mL EGF and FGF2. The mNSCs at the third generation were coated with 10ug/mL polylysine and induced to differentiate in the DMEM/F12 supplemented with 1% fetal bovine serum and 1% N2. MAIN OUTCOME MEASURES： The neural spheres of the third passage were identified by nestin immunofluorescence; at the same time, the cells were induced to differentiate, and the types of differentiated cell were identified by immunofluorescence for β Ⅲ tubulin, GFAP and CNPase. RESULTS： Seven days after primary culture, a great many neurospheres could be obtained by successive pasage. Immunofluorescence assays showed that the neurospheres were nestin positive, and after differentiation, the cells expressed GFAP, CNPase and β -Ⅲ-tu展开更多
β-mercaptoethanol induces in vitro adult adipose-derived stromal cells （ADSCs） to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the p...β-mercaptoethanol induces in vitro adult adipose-derived stromal cells （ADSCs） to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker （nestin）, a neuronal marker （neuron-specific enolase）, and a glial marker （glial fibrillary acidic protein）. In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.展开更多
Objective To review the in vitro development of bone marrow mesenchymal stem cells culture （BM-MSC）.Data sources The data cited in this review were mainly obtained from articles listed in Medline and PubMed. The s...Objective To review the in vitro development of bone marrow mesenchymal stem cells culture （BM-MSC）.Data sources The data cited in this review were mainly obtained from articles listed in Medline and PubMed. The search terms were “bone marrow mesenchymal stem cell” and “cell culture”.Study selection Articles regarding the in vitro development of BM-MSCs culture, as well as the challenge of optimizing cell culture environment in two-dimensional （2D） vs. 3D.Results Improving the culture conditions increases the proliferation and reduces the differentiation. Optimal values for many culture parameters remain to be identified. Expansion of BM-MSCs under defined conditions remains challenging, including the development of optimal culture conditions for BMSC and large-volume production systems.Conclusions Expansion of BM-MSCs under defined conditions remains challenges, including the development of optimal culture conditions for BMSC and scale-up to large-volume production systems. Optimal values for many culture parameters remain to be identified.展开更多
The human hepatitis B virus （HBV） and the duck hepatitis B virus （DHBV） share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and th...The human hepatitis B virus （HBV） and the duck hepatitis B virus （DHBV） share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and the coding open reading frames （ORFs） overlap extensively. In addition, the genomic and structural organization, as well as replication and biological characteristics, are very similar in both viruses. Host of the key features of hepadnaviral infection were first discovered in the DHBV model system and subsequently confirmed for HBV. There are, however, several differences between human HBV and DHBV. This review will focus on the molecular and cellular biology, evolution, and host adaptation of the avian hepatitis B viruses with particular emphasis on DHBV as a model system.展开更多
OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertili...OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.展开更多
AIMTo investigate whether the abnormal differentiation of the pterygium epithelium is related to the extracellular signal-regulated kinase (ERK) signaling pathway in vitro.METHODSThe expression levels of phosphorylate...AIMTo investigate whether the abnormal differentiation of the pterygium epithelium is related to the extracellular signal-regulated kinase (ERK) signaling pathway in vitro.METHODSThe expression levels of phosphorylated ERK (P-ERK), keratin family members including K19 and K10 and the ocular master control gene Pax-6 were measured in 16 surgically excised pterygium tissues and 12 eye bank conjunctiva. In colony-forming cell assays, the differences in clone morphology and in K10, K19, P-ERK and Pax-6 expression between the head and body were investigated. When cocultured with the ERK signaling pathway inhibitor PD98059, the changes in clone morphology, colony-forming efficiency, differentiated marker K10, K19 and Pax-6 expression and P-ERK protein expression level were examined by immunoreactivity and Western blot analysis.RESULTSThe expression of K19 and Pax-6 decreased in the pterygium, especially in the head. No staining of K10 was found in the normal conjunctiva epithelium, but it was found to be expressed in the superficial cells in the head of the pterygium. Characteristic upregulation of P-ERK was observed by immunohistochemistry. The clone from the head with more differentiated cells in the center expressed more K10, and the clone from the body expressed more K19. The P-ERK protein level increased in the pterygium epithelium compared with conjunctiva and decreased when cocultured with PD98059. The same medium with the ERK inhibitor PD98059 was more effective in promoting clonal growth than conventional medium with 3T3 murine feeder layers. It was observed that the epithelium clone co-cultured with the inhibitor had decreased K10 expression and increased K19 and Pax-6 expression.CONCLUSIONWe suggest ERK signaling pathway activation might play a role in the pterygium epithelium abnormal differentiation.展开更多
The studies from 7 or 8 days old mice were used to prepare the spermatogonial stem cells. The isolation and purification of spermatogonial stem cells were done by means of the Percoll discontinual density gradient con...The studies from 7 or 8 days old mice were used to prepare the spermatogonial stem cells. The isolation and purification of spermatogonial stem cells were done by means of the Percoll discontinual density gradient concentration. The cells of the 3rd band were collected and cultured in vitro in DMEM supplemented with 2.5% or 10% fetal bovine serum （FBS）. The results showed that with the feeder layer and 2.5% serum, the spermatogonial stem cells could proliferate, differentiate last more than 4 months.展开更多
Pluripotent stem cells(PSCs) are able to differentiate into several cell types, including pancreatic β cells. Differentiation of pancreatic β cells depends on certain transcription factors, which function in a coord...Pluripotent stem cells(PSCs) are able to differentiate into several cell types, including pancreatic β cells. Differentiation of pancreatic β cells depends on certain transcription factors, which function in a coordinated way during pancreas development. The existing protocols for in vitro differentiation produce pancreatic β cells, which are not highly responsive to glucose stimulation except after their transplantation into immune-compromised mice and allowing several weeks for further differentiation to ensure the maturation of these cells in vivo. Thus, although the substantial improvement that has been made for the differentiation of induced PSCs and embryonic stem cells toward pancreatic β cells, several challenges still hindering their full generation. Here, we summarize recent advances in the differentiation of PSCs into pancreatic β cells and discuss the challenges facing their differentiation as well as the different applications of these potential PSC-derived β cells.展开更多
SHANK2 is a scaffold protein that serves as a protein anchor at the postsynaptic density in neurons.Genetic variants of SHANK2 are strongly associated with synaptic dysfunction and the pathophysiology of autism spectr...SHANK2 is a scaffold protein that serves as a protein anchor at the postsynaptic density in neurons.Genetic variants of SHANK2 are strongly associated with synaptic dysfunction and the pathophysiology of autism spectrum disorder.Recent studies indicate that early neuronal developmental defects play a role in the pathogenesis of autism spectrum disorder,and that insulin-like growth factor 1 has a positive effect on neurite development.To investigate the effects of SHANK2 knockdown on early neuronal development,we generated a sparse culture system using human induced pluripotent stem cells,which then differentiated into neural progenitor cells after 3–14 days in culture,and which were dissociated into single neurons.Neurons in the experimental group were infected with shSHANK2 lentivirus carrying a red fluorescent protein reporter(shSHANK2 group).Control neurons were infected with scrambled sh Control lentivirus carrying a red fluorescent protein reporter(sh Control group).Neuronal somata and neurites were reconstructed based on the lentiviral red fluorescent protein signal.Developmental dendritic and motility changes in VGLUT1^+glutamatergic neurons and TH+dopaminergic neurons were then evaluated in both groups.Compared with sh Control VGLUT1^+neurons,the dendritic length and arborizations of shSHANK2 VGLUT1^+neurons were shorter and fewer,while cell soma speed was higher.Furthermore,dendritic length and arborization were significantly increased after insulin-like growth factor 1 treatment of shSHANK2 neurons,while cell soma speed remained unaffected.These results suggest that insulin-like growth factor 1 can rescue morphological defects,but not the change in neuronal motility.Collectively,our findings demonstrate that SHANK2 deficiency perturbs early neuronal development,and that IGF1 can partially rescue the neuronal defects caused by SHANK2 knockdown.All experimental procedures and protocols were approved by the Laboratory Animal Ethics Committee of Jinan University,China(approval No.20170228010)on February 2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
Mechanical stretch plays an important role in the control of cardiomyocyte behavior,as well as in the study of the mechanisms of cardiovascular function and pathology.The complexity involved in biological systems in v...Mechanical stretch plays an important role in the control of cardiomyocyte behavior,as well as in the study of the mechanisms of cardiovascular function and pathology.The complexity involved in biological systems in vivo has created a need for better in vitro techniques,thus a variety of cell stretching devices have been developed for a deeper understanding of cellular responses to strain.In this review,we introduce the design,functionality,and characteristics of multiple types of cell stretching devices from two and three dimensions,then discuss the research progress of promoting cardiomyogenic differentiation of stem cells by external stretching and its application in cardiac tissue engineering.展开更多
基金the National Natural Science Foundation of China,No.30672151
文摘BACKGROUND： Midbrain-derived neural stem cells （mNSCs） can differentiate into functional mature dopaminergic neurons. The mNSCs are considered the ideal choice for cell therapy of Parkinson＇s disease. OBJECTIVE： To isolate rat embryonic mNSCs and to observe the differentiation characteristics of mNSCs induced by cell growth-promoting factors. DESIGN, TIME AND SETTING： An in vitro cell culture study based on the molecular biology of nerve cells was carried out at the Institute of Clinical Medicine, China-Japan Friendship Hospital （China） from March to November 2007. MATERIALS： Sprague Dawley rats at embryonic day 14 were used in this study. Nestin antibody, β-Ⅲ tubulin antibody, glial fibrillary acidic protein （GFAP） antibody and cyclic nucleotide 3＇-phosphohydrolase （CNPase） antibody were provided by Abcam; DMEM/F12 medium and N2 supplement were provided by Invitrogen; epidermal growth factor （EGF） and fibroblast growth factor-2 （FGF2） were provided by R＆D Systems. METHODS： The ventral mesencephalon was dissected from embryonic day 14 rat embryos. By trypsin digestion and mechanical separation, the brain tissue was triturated into a fine single-cell suspension. The cells were cultured in 5 mL serum-free medium containing DMEM/FI 2, 1% N： supplement, 20 ng/mL EGF and FGF2. The mNSCs at the third generation were coated with 10ug/mL polylysine and induced to differentiate in the DMEM/F12 supplemented with 1% fetal bovine serum and 1% N2. MAIN OUTCOME MEASURES： The neural spheres of the third passage were identified by nestin immunofluorescence; at the same time, the cells were induced to differentiate, and the types of differentiated cell were identified by immunofluorescence for β Ⅲ tubulin, GFAP and CNPase. RESULTS： Seven days after primary culture, a great many neurospheres could be obtained by successive pasage. Immunofluorescence assays showed that the neurospheres were nestin positive, and after differentiation, the cells expressed GFAP, CNPase and β -Ⅲ-tu
文摘β-mercaptoethanol induces in vitro adult adipose-derived stromal cells （ADSCs） to differentiate into neurons. However, the ultrastructural features of the differentiated neuronal-like cells remain unknown. In the present study, inverted phase contrast microscopy was utilized to observe β-mercaptoethanol-induced differentiation of neuronal-like cells from human ADSCs, and immunocytochemistry and real-time polymerase chain reaction were employed to detect expression of a neural stem cells marker （nestin）, a neuronal marker （neuron-specific enolase）, and a glial marker （glial fibrillary acidic protein）. In addition, ultrastructure of neuronal-like cells was observed by transmission election microscopy. Results revealed highest expression rate of nestin and neuron-specific enolase at 3 and 5 hours following induced differentiation; cells in the 5-hour induction group exhibited a neuronal-specific structure, i.e., Nissl bodies. However, when induction solution was replaced by complete culture medium after 8-hour induction, the differentiated cells reverted to the fibroblast-like morphology from day 1. These results demonstrate that β-mercaptoethanol-induced ADSCs induced differentiation into neural stem cells, followed by morphology of neuronal-like cells. However, this differentiation state was not stable.
文摘Objective To review the in vitro development of bone marrow mesenchymal stem cells culture （BM-MSC）.Data sources The data cited in this review were mainly obtained from articles listed in Medline and PubMed. The search terms were “bone marrow mesenchymal stem cell” and “cell culture”.Study selection Articles regarding the in vitro development of BM-MSCs culture, as well as the challenge of optimizing cell culture environment in two-dimensional （2D） vs. 3D.Results Improving the culture conditions increases the proliferation and reduces the differentiation. Optimal values for many culture parameters remain to be identified. Expansion of BM-MSCs under defined conditions remains challenging, including the development of optimal culture conditions for BMSC and large-volume production systems.Conclusions Expansion of BM-MSCs under defined conditions remains challenges, including the development of optimal culture conditions for BMSC and scale-up to large-volume production systems. Optimal values for many culture parameters remain to be identified.
基金Supported by the Freie und Hansestadt Hamburg and the Bundesministcrium für Gesundheit und Soziale Sicherung grants from DFG and by the German Competence Network for Viral Hepatitis （Hop-Net）, funded by the German Ministry of Education and Research （BMBF）, Grant No. TFI3. IWe apologize to those authors whose work we could not cite directly due to space limitations. The authors are indebted to Claudia Franke （Heinrich-Pette-Institute, Hamburg, Germany） for providing the picture of core protein phosphorylation.
文摘The human hepatitis B virus （HBV） and the duck hepatitis B virus （DHBV） share several fundamental features. Both viruses have a partially double-stranded DNA genome that is replicated via a RNA intermediate and the coding open reading frames （ORFs） overlap extensively. In addition, the genomic and structural organization, as well as replication and biological characteristics, are very similar in both viruses. Host of the key features of hepadnaviral infection were first discovered in the DHBV model system and subsequently confirmed for HBV. There are, however, several differences between human HBV and DHBV. This review will focus on the molecular and cellular biology, evolution, and host adaptation of the avian hepatitis B viruses with particular emphasis on DHBV as a model system.
基金Supported by the National Natural Science Foundation of China(No.81173294)
文摘OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.
基金Supported by National Natural Science Foundation of China (No.81100649)
文摘AIMTo investigate whether the abnormal differentiation of the pterygium epithelium is related to the extracellular signal-regulated kinase (ERK) signaling pathway in vitro.METHODSThe expression levels of phosphorylated ERK (P-ERK), keratin family members including K19 and K10 and the ocular master control gene Pax-6 were measured in 16 surgically excised pterygium tissues and 12 eye bank conjunctiva. In colony-forming cell assays, the differences in clone morphology and in K10, K19, P-ERK and Pax-6 expression between the head and body were investigated. When cocultured with the ERK signaling pathway inhibitor PD98059, the changes in clone morphology, colony-forming efficiency, differentiated marker K10, K19 and Pax-6 expression and P-ERK protein expression level were examined by immunoreactivity and Western blot analysis.RESULTSThe expression of K19 and Pax-6 decreased in the pterygium, especially in the head. No staining of K10 was found in the normal conjunctiva epithelium, but it was found to be expressed in the superficial cells in the head of the pterygium. Characteristic upregulation of P-ERK was observed by immunohistochemistry. The clone from the head with more differentiated cells in the center expressed more K10, and the clone from the body expressed more K19. The P-ERK protein level increased in the pterygium epithelium compared with conjunctiva and decreased when cocultured with PD98059. The same medium with the ERK inhibitor PD98059 was more effective in promoting clonal growth than conventional medium with 3T3 murine feeder layers. It was observed that the epithelium clone co-cultured with the inhibitor had decreased K10 expression and increased K19 and Pax-6 expression.CONCLUSIONWe suggest ERK signaling pathway activation might play a role in the pterygium epithelium abnormal differentiation.
文摘The studies from 7 or 8 days old mice were used to prepare the spermatogonial stem cells. The isolation and purification of spermatogonial stem cells were done by means of the Percoll discontinual density gradient concentration. The cells of the 3rd band were collected and cultured in vitro in DMEM supplemented with 2.5% or 10% fetal bovine serum （FBS）. The results showed that with the feeder layer and 2.5% serum, the spermatogonial stem cells could proliferate, differentiate last more than 4 months.
文摘Pluripotent stem cells(PSCs) are able to differentiate into several cell types, including pancreatic β cells. Differentiation of pancreatic β cells depends on certain transcription factors, which function in a coordinated way during pancreas development. The existing protocols for in vitro differentiation produce pancreatic β cells, which are not highly responsive to glucose stimulation except after their transplantation into immune-compromised mice and allowing several weeks for further differentiation to ensure the maturation of these cells in vivo. Thus, although the substantial improvement that has been made for the differentiation of induced PSCs and embryonic stem cells toward pancreatic β cells, several challenges still hindering their full generation. Here, we summarize recent advances in the differentiation of PSCs into pancreatic β cells and discuss the challenges facing their differentiation as well as the different applications of these potential PSC-derived β cells.
基金supported by the National Natural Science Foundation of China,No.81771222(to LLS)the Natural Science Foundation of Guangdong Province of China,No.2019A1515011316(to LLS)+2 种基金the National Key Research and Development Program of China,Stem Cell and Translational Research,No.2017YFA0105102(to LLS)the Guangzhou Science and Technology Innovation Development Special Fund Project of China,No.201804010212(to LLS)the Program of Introducing Talents of Discipline to Universities of China,No.B14036(to KFS)。
文摘SHANK2 is a scaffold protein that serves as a protein anchor at the postsynaptic density in neurons.Genetic variants of SHANK2 are strongly associated with synaptic dysfunction and the pathophysiology of autism spectrum disorder.Recent studies indicate that early neuronal developmental defects play a role in the pathogenesis of autism spectrum disorder,and that insulin-like growth factor 1 has a positive effect on neurite development.To investigate the effects of SHANK2 knockdown on early neuronal development,we generated a sparse culture system using human induced pluripotent stem cells,which then differentiated into neural progenitor cells after 3–14 days in culture,and which were dissociated into single neurons.Neurons in the experimental group were infected with shSHANK2 lentivirus carrying a red fluorescent protein reporter(shSHANK2 group).Control neurons were infected with scrambled sh Control lentivirus carrying a red fluorescent protein reporter(sh Control group).Neuronal somata and neurites were reconstructed based on the lentiviral red fluorescent protein signal.Developmental dendritic and motility changes in VGLUT1^+glutamatergic neurons and TH+dopaminergic neurons were then evaluated in both groups.Compared with sh Control VGLUT1^+neurons,the dendritic length and arborizations of shSHANK2 VGLUT1^+neurons were shorter and fewer,while cell soma speed was higher.Furthermore,dendritic length and arborization were significantly increased after insulin-like growth factor 1 treatment of shSHANK2 neurons,while cell soma speed remained unaffected.These results suggest that insulin-like growth factor 1 can rescue morphological defects,but not the change in neuronal motility.Collectively,our findings demonstrate that SHANK2 deficiency perturbs early neuronal development,and that IGF1 can partially rescue the neuronal defects caused by SHANK2 knockdown.All experimental procedures and protocols were approved by the Laboratory Animal Ethics Committee of Jinan University,China(approval No.20170228010)on February 2
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This work was supported by the National Natural Science Foundation of China(Nos.U20A20390,11827803,and 11302020)the 111 Project(No.B13003).
文摘Mechanical stretch plays an important role in the control of cardiomyocyte behavior,as well as in the study of the mechanisms of cardiovascular function and pathology.The complexity involved in biological systems in vivo has created a need for better in vitro techniques,thus a variety of cell stretching devices have been developed for a deeper understanding of cellular responses to strain.In this review,we introduce the design,functionality,and characteristics of multiple types of cell stretching devices from two and three dimensions,then discuss the research progress of promoting cardiomyogenic differentiation of stem cells by external stretching and its application in cardiac tissue engineering.
