中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (50): 8168-8173.doi: 10.3969/j.issn.2095-4344.2014.50.023

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

人胚胎脑组织蛋白提取物诱导人胎盘间充质干细胞向神经细胞的分化

马丽君1，范  恒1，马晓娜1，魏  军1，2，李玉奎1，2   

  1. 1宁夏医科大学总医院干细胞研究所，宁夏回族自治区银川市  750004；2宁夏医科大学生育力保持省部共建教育部重点实验室，宁夏回族自治区银川市  750004
  • 出版日期:2014-12-03 发布日期:2014-12-03
  • 通讯作者: 魏军，硕士，教授，宁夏医科大学总医院干细胞研究所，宁夏回族自治区银川市750004；宁夏医科大学生育力保持省部共建教育部重点实验室，宁夏回族自治区银川市 750004 并列通讯作者：李玉奎，博士，教授，宁夏医科大学总医院干细胞研究所，宁夏回族自治区银川市 750004；宁夏医科大学生育力保持省部共建教育部重点实验室，宁夏回族自治区银川市 750004
  • 作者简介:马丽君，女，1981年生，山东省济南市人，回族，在读博士，助理研究员，主要从事干细胞生物学研究。
  • 基金资助:

    宁夏自然科学基金项目(NZ1229)；宁夏医科大学校级课题(XM201135)

Protein extracts from human embryonic brain tissue induce the neuronal differentiation of mesenchymal stem cells from human placenta

Ma Li-jun1, Fan Heng1, Ma Xiao-na1, Wei Jun1, 2, Li Yu-kui1, 2   

  1. 1Ningxia Human Stem Cell Institute, the General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 2Province-Ministry Co-Constructed Key Laboratory of Fecundity Preserve, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Online:2014-12-03 Published:2014-12-03
  • Contact: Wei Jun, Master, Professor, Ningxia Human Stem Cell Institute, the General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Province-Ministry Co-Constructed Key Laboratory of Fecundity Preserve, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Corresponding author: Li Yu-kui, M.D., Professor, Ningxia Human Stem Cell Institute, the General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Province-Ministry Co-Constructed Key Laboratory of Fecundity Preserve, Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • About author:Ma Li-jun, Studying for doctorate, Assistant researcher, Ningxia Human Stem Cell Institute, the General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:

    the Natural Science Foundation of Ningxia Hui Autonomous Region, No. NZ1229; the Project of Ningxia Medical University, No. XM201135

PDF

394

被引次数

1

摘要：

背景：选择合适的诱导方法诱导人胎盘间充质干细胞分化为神经元样细胞是临床治疗神经损伤的关键。
目的：在人胎盘间充质干细胞中加入商品化的人胚胎脑组织蛋白提取物，观察其诱导分化为神经元样细胞的可行性。
方法：采用酶消化法分离培养人胎盘间充质干细胞，传至第3代，将人早期胚胎脑组织蛋白提取物加入诱导培养基培养6 d，加入浓度为20 nmol/L全反式维甲酸和500 μg/L音猬因子培养3 d，换新的培养液，包含体积分数为10%胎牛血清，10 μg/L脑源性神经营养因子，20 μg/L神经胶质细胞源性神经营养因子，50 μg/L胰岛素样生长因子Ⅱ型继续培养3 d。
结果与结论：胎盘组织经酶消化后获得贴壁细胞，传至第2代细胞形态为梭形，成漩涡样生长，传至第3代细胞形态较均一。诱导后细胞经免疫细胞化学法检测均表达神经元特异性烯醇化酶、巢蛋白、胶质纤维酸性蛋白、神经丝蛋白、神经生长相关蛋白43；ELISA法检测细胞培养上清脑源性神经营养因子、神经营养因子3、神经营养因子4为阳性表达；RT-PCR检测细胞微管相关蛋白、神经元特异性烯醇化酶、神经丝蛋白、神经生长相关蛋白43均为阳性表达。结果表明人胚胎脑组织蛋白提取物使人胎盘间充质干细胞诱导分化为神经元样细胞。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

关键词: 干细胞, 诱导, 人胎盘间充质干细胞, 人早期胚胎脑组织蛋白提取物, 神经细胞, 分化, 宁夏自然科学基金

Abstract:

BACKGROUND: It is a key to choose an appropriate method to trans-differentiate mesenchymal stem cells from human placenta into neuron-like cells for clinical treatment of neural system injury.
OBJECTIVE: To observe the feasibility of the neuronal differentiation of mesenchymal stem cells from human placenta with protein extracts of human embryonic brain tissue.
METHODS: Mesenchymal stem cells from human placenta were isolated and cultured using enzyme digestion method. The third passage of cells were incubated in induction medium containing protein extracts of human embryonic brain tissue for 6 days, and then cultured in culture medium containing 20 nmol/L all-trans retinoic acid and 500 μg/L sonic hedgehog for 3 days, followed by 3-day continuous culture in 10 μg/L brain-derived neurotrophic factor, 20 μg/L glial cell line-derived neurotrophic factor, 50 μg/L insulin-like growth factor type II
RESULTS AND CONCLUSION: The adherent cells were obtained after enzyme digestion of placenta tissue. The passage 2 cells were fusiform-shaped and exhibited a whirlpool-like growth. The passage 3 cells were uniform in 
morphology. After induction, immunocytochemistry assay showed that the cells expressed neuron-specific enolase, nestin, glial fibrillary acidic protein, neurofilament protein, and nerve growth associated protein 43; ELISA results showed that cells were positive for brain-derived neurotrophic factor, neurotrophin 3, neurotrophin 4; RT-PCR results showed that cells were positive for microtubule-associated protein, neuron-specific enolase, neurofilament protein, nerve growth associated protein 43. Results indicate that mesenchymal stem cells from human placenta can differentiate into neuron-like cells under induction of protein extracts of human embryonic brain tissue.


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程


全文链接：

Key words: placenta, mesenchymal stem cells, neurons, embryonic induction, cell differentiation

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