中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5944-5948.doi: 10.3969/j.issn.1673-8225.2010.32.012

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

酶消化和整体灌注分离人胎盘间充质干细胞的比较

李 芳1，2，苗宗宁3，许云云4 ，张学光1，2   

  1. 1苏州大学基础医学与生物科学学院人体解剖与组织胚胎系，江苏省苏州市  215007；2江苏省干细胞重点实验室，江苏省苏州市215007；3无锡市第三人民医院烧伤研究所，江苏省无锡市214041；4苏州大学附属儿童医院儿科研究所，江苏省苏州市215007
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 张学光，教授，博士生导师，江苏省干细胞研究重点实验室，江苏省苏州市 215007 xueguangzh@ yahoo.com.cn
  • 作者简介:李芳☆，女，1979年生，陕西省西安市人，汉族，苏州大学基础医学与生物科学学院在读博士，讲师，主要从事人胚胎生殖干细胞，胎盘间充质干细胞的研究。 mercuryfish@ 163.com

Isolation of human placenta derived mesenchymal stem cells by enzymatic digestion versus global perfusion

Li Fang 1,2 , Miao Zong-ning3, Xu Yun-yun4, Zhang Xue-guang 1,2   

  1. 1 Department of Human Anatomy, Histology and Embryology, College of Basic Medicine and Biological Science, Soochow University, Suzhou  215007, Jiangsu Province, China; 2 Jiangsu Stem Cell Key Laboratory, Suzhou  215007, Jiangsu Province, China; 3 Burn Institute, Wuxi Third People’s Hospital, Wuxi  214041, Jiangsu Province, China; 4 Institute of Pediatrics, Children Hospital Affiliated to Soochow University, Suzhou  215007, Jiangsu Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Zhang Xue-guang, Professor, Doctoral supervisor, Department of Human Anatomy, Histology and Embryology, College of Basic Medicine and Biological Science, Soochow University, Suzhou 215007, Jiangsu Province, China; Jiangsu Stem Cell Key Laboratory, Suzhou 215007, Jiangsu Province, China xueguangzh@yahoo.com.cn
  • About author:Li Fang☆, Studying for doctorate, Lecturer, Department of Human Anatomy, Histology and Embryology, College of Basic Medicine and Biological Science, Soochow University, Suzhou 215007, Jiangsu Province, China; Jiangsu Stem Cell Key Laboratory, Suzhou 215007, Jiangsu Province, China mercuryfish@163.com

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337

被引次数

8

摘要：

背景：目前，分离和纯化人胎盘间充质干细胞的方法带有一定的盲目性，因为胎盘组织中其他细胞的干扰情况严重。
目的：探讨酶消化和整体灌注的方法分离培养人胎盘间充质干细胞的效果，寻找一种高效的分离方法，以建立稳定的体外培养体系。
方法：取完整的胎盘组织，按照不同的培养方法分离人胎盘间充质干细胞，酶消化组采用Ⅳ型胶原酶消化分离；整体灌注组采用整体灌注的方法进行分离。常规培养后对比两种方法分离培养细胞的生长特性、表面抗原表达以及多向分化潜能。
结果及结论：两种分离方法培养的细胞在生长特性、多向分化潜能方面没有明显区别，流式细胞检测酶消化组细胞表达CD90明显低于整体灌注组细胞(P < 0.05)。可见酶消化和整体灌注的方法都可以分离人胎盘间充质干细胞，整体灌注的方法分离纯度更高。

关键词: 整体灌注, 胎盘间充质干细胞, 胶原酶, 分离, 培养

Abstract:

BACKGROUND: The method of isolation and purification of human placenta derived mesenchymal stem cells (PMSCs) has certain blindness due to the interference of other cells within the placenta.
OBJECTIVE: To investigate the efficiency of two methods to isolate PMSCs such as enzyme digestion and global perfusion, to find an efficient method and to establish a stable in vitro culture system of PMSCs.
METHODS: The normal term placenta was obtained to isolate PMSCs by different methods. Ⅳ collagenase digestion was used in the enzymatic digestion group, and global perfusion was utilized in the global perfusion group. The growth characteristics of cells, cell surface marker, as well as the multiple differentiation potential of the cultured cell obtain by these two methods were compared following conventional culture.
RESULTS AND CONCLUSION: There was no significant difference of the cell growth, multiple differentiation potential between the cultured cell obtain by these two method mentioned above, but the CD90 was obviously lowly expressed on the cultured cells obtained by enzymatic digestion compared with global perfusion (P < 0.05), which shows that global perfusion is a more efficient way to get higher purity of PMSCs compared with enzymatic digestion.

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