中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5939-5943.doi: 10.3969/j.issn.1673-8225.2010.32.011

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

体外羊膜间充质干细胞分离培养及向神经元样细胞的分化

金 钧，黄 坚，王 俊，付建红   

  1. 苏州大学附属第一医院急诊科，江苏省苏州市   215006
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 作者简介:金钧★，男，1970年生，江苏省苏州市人，汉族，1999年苏州医学院毕业，硕士，副主任医师，主要从事危重病医学临床治疗和研究。stemcell2004@ 163.com

Isolation, culture and differentiation of amnion mesenchymal stem cells into neuron-like cells in vitro 

Jin Jun, Huang Jian, Wang Jun, Fu Jian-hong   

  1. Department of Emergency, First Affiliated Hospital, Soochow University, Suzhou  215006, Jiangsu Province, China
  • Online:2010-08-06 Published:2010-08-06
  • About author:Jin Jun★, Master, Associate chief physician, Department of Emergency, First Affiliated Hospital, Soochow University, Suzhou 215006, Jiangsu Province, China stemcell2004@163. com

PDF

409

被引次数

16

摘要：

背景：胎盘组织作为寻找人类间充质干细胞的新来源，已成为备受人们关注的研究热点。从胎盘组织中羊膜层分离、培养细胞，因其来源广泛、不受伦理限制等优越性而具有广阔的应用前景。
目的：观察人羊膜间充质干细胞体外分离培养的方法，及向神经元样细胞分化的能力。
方法：用酶消化法从羊膜中分离和培养间充质干细胞，并通过形态的均一性及流式细胞术检测其表面标志以鉴定纯度。应用DMEM-LG+20 g/L DMSO+100 µmol/L BHA+10 μg/L碱性成纤维细胞生长因子诱导分化，通过免疫荧光染色鉴定诱导后的细胞。
结果与结论：可从羊膜组织成功分离培养出间充质干细胞，细胞贴壁生长，在体外短期内可以稳定增殖和传代。具有与骨髓间充质干细胞相似的表面标志，体外可以诱导分化为神经元样细胞，胶质纤维酸性蛋白与神经元特异性烯醇化酶免疫荧光染色均为阳性。提示羊膜间充质干细胞可以在体外分离培养并诱导分化为神经元样细胞，羊膜组织可以作为干细胞研究以及神经组织疾病细胞治疗的一个全新的细胞来源。

关键词: 羊膜, 间充质干细胞, 细胞培养, 诱导分化, 神经元

Abstract:

BACKGROUND: Placenta as a new source of seeking mesenchymal stem cells has been paid great attention. To harvest cells from amnion layer of placenta has wide application prospects due to its advantages, such as extensive source and no ethical limitation.
OBJECTIVE: To study the method of in vitro isolating human amnion mesenchymal stem cells, and the ability of differentiating into neuron-like cells.
METHODS: Mesenchymal stem cells were harvested and cultured from amnion by enzyme digestion method. The surface marker and purity of human amnion mesenchymal stem cells were measured by uniform morphology and flow cytometry. The human amnion mesenchymal stem cells were induced to differentiate using DMEM-LG+20 g/L DMSO+100 µmol/L BHA+ 10 μg/L basic fibroblast growth factor. The differentiated cells were identified with immunofluorescence staining.
RESULTS AND CONCLUSION: Mesenchymal stem cells could be successfully harvested from amnion. Cells grew and adhered to the wall, and could proliferate and subculture in a short period in vitro. Amnion mesenchymal stem cells had similar surface markers to bone marrow mesenchymal stem cells, and could differentiate into neuron-like cells in vitro. Amnion mesenchymal stem cells were positive for glial fibrillary acid protein and neuron specific enolase. Results have suggested that amnion mesenchymal stem cells could be induced to differentiate into neuron-like cells and amnion tissue could be a novel source in stem cells study and for nerve tissue diseases.

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