BACKGROUND： It has long been thought that neurons and glial cells are produced from distinct progenitor pools, but recent studies suggest that the glial progenitor cell in the subventricular zone can generate neurons...BACKGROUND： It has long been thought that neurons and glial cells are produced from distinct progenitor pools, but recent studies suggest that the glial progenitor cell in the subventricular zone can generate neurons in the adult rodent brain. OBJECTIVE： To investigate the ability of a telomerase-immortalized human glial progenitor cell line to differentiate into functionally mature neurons. DESIGN, TIME AND SETTING： The cellular and molecular biology experiment was performed at the Cell Biology Laboratory in the School of Basic Medical Sciences, Peking University Health Science Center, between July 2007 and May 2008. MATERIALS： A telomerase reverse transcriptase immortalized human glial progenitor cell line, was established in our laboratory. Dibutyryl cyclic AMP was purchased from Sigma （USA）. Specific antibodies against glial fibrillary acidic protein, ~ -tubulin-Ⅲand A2B5 were purchased from Chemicon, USA. Polyclonal antibodies against nestin and MAP2ab were obtained from Neomarker, USA. METHODS： The telomerase immortalized human glial progenitor cell line was passaged and maintained in growth medium consisting of DMEM/F12 （1：1） with N2 supplement （1%, v/v）, L-Glutamine （2 mmol/L）, epidermal growth factor （20 ng/mL）, basic fibroblast growth factor （20 ng/mL） and 3% fetal bovine serum. Neuronal differentiation was induced by the addition of 1 mmol/L dibutyryl cyclic AMP and 10% fetal bovine serum. MAIN OUTCOME MEASURES： Neuronal differentiation was evaluated by RT-PCR, quantitative PCR, immunofluorescence staining, Western blot analysis and electrophysiology. RESULTS： After dibutyryl cyclic AMP induction in the telomerase immortalized human glial progenitor cells, the expression of neuronal-specific marker mRNAs and proteins increased significantly. Concurrently, an apparent fast inward Na^＋ current was evoked in the cells after induction. CONCLUSION： This study suggests that some human glial progenitor ceils are indeed capable of generating functionally mature neurons, and such cells may be useful for treating human neurological disorders.展开更多
Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes （microtubule-associated protein-2, glial fibrillary ...Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes （microtubule-associated protein-2, glial fibrillary acidic protein and nestin） in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1 2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.展开更多
The peripheral nerve injuries,representing some of the most common types of traumatic lesions affecting the nervous system,are highly invalidating for the patients besides being a huge social burden.Although periphera...The peripheral nerve injuries,representing some of the most common types of traumatic lesions affecting the nervous system,are highly invalidating for the patients besides being a huge social burden.Although peripheral nervous system owns a higher regenerative capacity than does central nervous system,mostly depending on Schwann cells intervention in injury repair,several factors determine the extent of functional outcome after healing.Based on the injury type,different therapeutic approaches have been investigated so far.Nerve grafting and Schwann cell transplantation have represented the gold standard treatment for peripheral nerve injuries,however these approaches own limitations,such as scarce donor nerve availability and donor site morbidity.Cell based therapies might provide a suitable tool for peripheral nerve regeneration,in fact,the ability of different stem cell types to differentiate towards Schwann cells in combination with the use of different scaffolds have been widely investigated in animal models of peripheral nerve injuries in the last decade.Dental pulp is a promising cell source for regenerative medicine,because of the ease of isolation procedures,stem cell proliferation and multipotency abilities,which are due to the embryological origin from neural crest.In this article we review the literature concerning the application of tooth derived stem cell populations combined with different conduits to peripheral nerve injuries animal models,highlighting their regenerative contribution exerted through either glial differentiation and neuroprotective/neurotrophic effects on the host tissue.展开更多
Background There are definite gender differences in patients with macular holes. Menopausal women over 50 years are most affected. We aimed to observe the effect of estrogen on collagen gel contraction by cultured hum...Background There are definite gender differences in patients with macular holes. Menopausal women over 50 years are most affected. We aimed to observe the effect of estrogen on collagen gel contraction by cultured human retinal glial cells. It is speculated that estrogen could strengthen the tensile stress of the macula by maintaining the correct morphology and contraction. Methods Estrogen was used to determine its effects on collagen gel contraction, and its function was measured using morphological changes in cells. Human retinal glial cells were cultured in collagen solution. The cells were then exposed to collagen gels and the degree of contraction of the gel was determined. Results Estrogen at differing concentrations had no effect on the growth of human retinal glial cells. However, after exposed to collagen gel block, less contraction was noted in the estrogen-treated group than in the control group. Conclusions Estrogen can inhibit collagen gel contraction by glial cells. These results suggest a mechanism for macular hole formation, which is observed in menopausal females.展开更多
背景:人尿源性干细胞是近年来新发现的成体干细胞,其来源不受限制,提取简便,具备良好的增殖能力及多向分化潜能,近年来已应用于泌尿系疾病中的神经功能修复,如应激性尿失禁以及膀胱输尿管返流等。目的:探索人尿源性干细胞向神经元样细...背景:人尿源性干细胞是近年来新发现的成体干细胞,其来源不受限制,提取简便,具备良好的增殖能力及多向分化潜能,近年来已应用于泌尿系疾病中的神经功能修复,如应激性尿失禁以及膀胱输尿管返流等。目的:探索人尿源性干细胞向神经元样细胞诱导分化能力及对大鼠脊髓损伤的修复作用。方法:体外获取人尿源性干细胞后利用流式细胞仪检测其细胞表型,将人尿源性干细胞向神经元样细胞诱导后进行免疫组织化学染色鉴定。采用Allen方法制作大鼠T9节段脊髓损伤模型,24只SD大鼠被随机分为2组:脊髓损伤组和人尿源性干细胞组,每组12只。人尿源性干细胞组在脊髓损伤后第1天于损伤脊髓边缘注入2μL细胞浓度为1.0×10^11L^-1人尿源性干细胞,脊髓损伤组注入等量含体积分数为10%胎牛血清的L-DMEM培养液,于造模后第1,10,20,30天进行BBB评分,第30天取各组损伤脊髓组织分别进行Luxol Fast Blue染色、小胶质细胞/巨噬细胞染色和胶质纤维酸性蛋白染色,并计算损伤脊髓面积和胶质纤维酸性蛋白荧光强度。结果与结论:①人尿源性干细胞高表达CD29、CD90,而低表达CD45,并且在体外可向神经元样细胞诱导分化;②造模后第1,10天两组大鼠BBB评分差异无显著性意义(P>0.05),第20,30天人尿源性干细胞组BBB评分明显高于脊髓损伤组(P<0.05);③人尿源性干细胞组脊髓损伤面积明显低于脊髓损伤组(P<0.05),胶质纤维酸性蛋白染色显示人尿源性干细胞组荧光强度明显低于脊髓损伤组(P<0.05);④结果表明,人尿源性干细胞能向神经元样细胞分化,并对大鼠脊髓损伤具有修复作用。展开更多
Following spinal cord injury, astrocyte proliferation and scar formation are the main factors inhibiting the regeneration and growth of spinal cord axons. Recombinant decorin suppresses inflammatory reactions, inhibit...Following spinal cord injury, astrocyte proliferation and scar formation are the main factors inhibiting the regeneration and growth of spinal cord axons. Recombinant decorin suppresses inflammatory reactions, inhibits glial scar formation, and promotes axonal growth. Rat models of T8 spinal cord contusion were created with the NYU impactor and these models were subjected to combined transplantation of bone morphogenetic protein-4-induced glial-restricted precursor-derived astro- cytes and human recombinant decorin transplantation. At 28 days after spinal cord contusion, dou- ble-immunofluorescent histochemistry revealed that combined transplantation inhibited the early in- flammatory response in injured rats. Furthermore, brain-derived neurotrophic factor, which was se- creted by transplanted cells, protected injured axons. The combined transplantation promoted ax- onal regeneration and growth of injured motor and sensory neurons by inhibiting astrocyte prolif- eration and glial scar formation, with astrocytes forming a linear arrangement in the contused spinal cord, thus providing axonal regeneration channels.展开更多
基金Supported by:the National Natural Science Foundation of China,No.30600167
文摘BACKGROUND： It has long been thought that neurons and glial cells are produced from distinct progenitor pools, but recent studies suggest that the glial progenitor cell in the subventricular zone can generate neurons in the adult rodent brain. OBJECTIVE： To investigate the ability of a telomerase-immortalized human glial progenitor cell line to differentiate into functionally mature neurons. DESIGN, TIME AND SETTING： The cellular and molecular biology experiment was performed at the Cell Biology Laboratory in the School of Basic Medical Sciences, Peking University Health Science Center, between July 2007 and May 2008. MATERIALS： A telomerase reverse transcriptase immortalized human glial progenitor cell line, was established in our laboratory. Dibutyryl cyclic AMP was purchased from Sigma （USA）. Specific antibodies against glial fibrillary acidic protein, ~ -tubulin-Ⅲand A2B5 were purchased from Chemicon, USA. Polyclonal antibodies against nestin and MAP2ab were obtained from Neomarker, USA. METHODS： The telomerase immortalized human glial progenitor cell line was passaged and maintained in growth medium consisting of DMEM/F12 （1：1） with N2 supplement （1%, v/v）, L-Glutamine （2 mmol/L）, epidermal growth factor （20 ng/mL）, basic fibroblast growth factor （20 ng/mL） and 3% fetal bovine serum. Neuronal differentiation was induced by the addition of 1 mmol/L dibutyryl cyclic AMP and 10% fetal bovine serum. MAIN OUTCOME MEASURES： Neuronal differentiation was evaluated by RT-PCR, quantitative PCR, immunofluorescence staining, Western blot analysis and electrophysiology. RESULTS： After dibutyryl cyclic AMP induction in the telomerase immortalized human glial progenitor cells, the expression of neuronal-specific marker mRNAs and proteins increased significantly. Concurrently, an apparent fast inward Na^＋ current was evoked in the cells after induction. CONCLUSION： This study suggests that some human glial progenitor ceils are indeed capable of generating functionally mature neurons, and such cells may be useful for treating human neurological disorders.
基金sponsored by the National Natural Science Foundation of China, No. 30271325the Natural Science Foundation of Jiangsu Province, No. BK2001170the National Basic Research Program of China (973 Program), No.2005CB522604
文摘Human amniotic epithelial cells were isolated from a piece of fresh amnion. Using immunocytochemical methods, we investigated the expression of neuronal phenotypes （microtubule-associated protein-2, glial fibrillary acidic protein and nestin） in human amniotic epithelial cells. The conditioned medium of human amniotic epithelial cells promoted the growth and proliferation of rat glial cells cultured in vitro, and this effect was dose-dependent. Human amniotic epithelial cells were further transplanted into the corpus striatum of healthy adult rats and the grafted cells could integrate with the host and migrate 1 2 mm along the nerve fibers in corpus callosum. Our experimental findings indicate that human amniotic epithelial cells may be a new kind of seed cells for use in neurograft.
文摘The peripheral nerve injuries,representing some of the most common types of traumatic lesions affecting the nervous system,are highly invalidating for the patients besides being a huge social burden.Although peripheral nervous system owns a higher regenerative capacity than does central nervous system,mostly depending on Schwann cells intervention in injury repair,several factors determine the extent of functional outcome after healing.Based on the injury type,different therapeutic approaches have been investigated so far.Nerve grafting and Schwann cell transplantation have represented the gold standard treatment for peripheral nerve injuries,however these approaches own limitations,such as scarce donor nerve availability and donor site morbidity.Cell based therapies might provide a suitable tool for peripheral nerve regeneration,in fact,the ability of different stem cell types to differentiate towards Schwann cells in combination with the use of different scaffolds have been widely investigated in animal models of peripheral nerve injuries in the last decade.Dental pulp is a promising cell source for regenerative medicine,because of the ease of isolation procedures,stem cell proliferation and multipotency abilities,which are due to the embryological origin from neural crest.In this article we review the literature concerning the application of tooth derived stem cell populations combined with different conduits to peripheral nerve injuries animal models,highlighting their regenerative contribution exerted through either glial differentiation and neuroprotective/neurotrophic effects on the host tissue.
文摘Background There are definite gender differences in patients with macular holes. Menopausal women over 50 years are most affected. We aimed to observe the effect of estrogen on collagen gel contraction by cultured human retinal glial cells. It is speculated that estrogen could strengthen the tensile stress of the macula by maintaining the correct morphology and contraction. Methods Estrogen was used to determine its effects on collagen gel contraction, and its function was measured using morphological changes in cells. Human retinal glial cells were cultured in collagen solution. The cells were then exposed to collagen gels and the degree of contraction of the gel was determined. Results Estrogen at differing concentrations had no effect on the growth of human retinal glial cells. However, after exposed to collagen gel block, less contraction was noted in the estrogen-treated group than in the control group. Conclusions Estrogen can inhibit collagen gel contraction by glial cells. These results suggest a mechanism for macular hole formation, which is observed in menopausal females.
文摘背景:人尿源性干细胞是近年来新发现的成体干细胞,其来源不受限制,提取简便,具备良好的增殖能力及多向分化潜能,近年来已应用于泌尿系疾病中的神经功能修复,如应激性尿失禁以及膀胱输尿管返流等。目的:探索人尿源性干细胞向神经元样细胞诱导分化能力及对大鼠脊髓损伤的修复作用。方法:体外获取人尿源性干细胞后利用流式细胞仪检测其细胞表型,将人尿源性干细胞向神经元样细胞诱导后进行免疫组织化学染色鉴定。采用Allen方法制作大鼠T9节段脊髓损伤模型,24只SD大鼠被随机分为2组:脊髓损伤组和人尿源性干细胞组,每组12只。人尿源性干细胞组在脊髓损伤后第1天于损伤脊髓边缘注入2μL细胞浓度为1.0×10^11L^-1人尿源性干细胞,脊髓损伤组注入等量含体积分数为10%胎牛血清的L-DMEM培养液,于造模后第1,10,20,30天进行BBB评分,第30天取各组损伤脊髓组织分别进行Luxol Fast Blue染色、小胶质细胞/巨噬细胞染色和胶质纤维酸性蛋白染色,并计算损伤脊髓面积和胶质纤维酸性蛋白荧光强度。结果与结论:①人尿源性干细胞高表达CD29、CD90,而低表达CD45,并且在体外可向神经元样细胞诱导分化;②造模后第1,10天两组大鼠BBB评分差异无显著性意义(P>0.05),第20,30天人尿源性干细胞组BBB评分明显高于脊髓损伤组(P<0.05);③人尿源性干细胞组脊髓损伤面积明显低于脊髓损伤组(P<0.05),胶质纤维酸性蛋白染色显示人尿源性干细胞组荧光强度明显低于脊髓损伤组(P<0.05);④结果表明,人尿源性干细胞能向神经元样细胞分化,并对大鼠脊髓损伤具有修复作用。
基金supported by funding from the Ministry of Finance People’s Republic of ChinaChina Rehabilitation Research Center Research Program grants, No. 2008-2,2008-3, 2008-4, 2008-5
文摘Following spinal cord injury, astrocyte proliferation and scar formation are the main factors inhibiting the regeneration and growth of spinal cord axons. Recombinant decorin suppresses inflammatory reactions, inhibits glial scar formation, and promotes axonal growth. Rat models of T8 spinal cord contusion were created with the NYU impactor and these models were subjected to combined transplantation of bone morphogenetic protein-4-induced glial-restricted precursor-derived astro- cytes and human recombinant decorin transplantation. At 28 days after spinal cord contusion, dou- ble-immunofluorescent histochemistry revealed that combined transplantation inhibited the early in- flammatory response in injured rats. Furthermore, brain-derived neurotrophic factor, which was se- creted by transplanted cells, protected injured axons. The combined transplantation promoted ax- onal regeneration and growth of injured motor and sensory neurons by inhibiting astrocyte prolif- eration and glial scar formation, with astrocytes forming a linear arrangement in the contused spinal cord, thus providing axonal regeneration channels.
