中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (32): 5891-5895.doi: 10.3969/j.issn.1673-8225.2011.32.001

• 骨髓干细胞 bone marrow stem cells •    下一篇

大鼠骨髓间充质干细胞培养的首次换液模式

倪  萍1，陈自谦2，彭德新2，白  伟2   

  1. 解放军南京军区福州总医院，1医学工程科，2医学影像中心，福建省福州市  350025
  • 收稿日期:2011-02-12 修回日期:2011-03-18 出版日期:2011-08-06 发布日期:2011-08-06
  • 通讯作者: 陈自谦，主任医师，教授，博士生导师，解放军南京军区福州总医院医学影像中心，福建省福州市 350025 chenziqianfz@yahoo.con.cn
  • 作者简介:倪萍★，女，1965年生，福建省福州市人，汉族，1990年福州大学毕业，硕士，高级工程师，主要从事细胞移植磁共振活体示踪成像研究。 nping6@sohu.com
  • 基金资助:

    福建省社会发展计划重点项目(2009Y0037)，福建省自然科学基金(2009J001187)，南京军区医学科技创新重点课题(09Z029)。

Modalities of the first medium exchange during the culture of rat bone-marrow derived mesenchymal stem cells

Ni Ping1, Chen Zi-qian2, Peng De-xin2, Bai Wei2   

  1. 1Department of Medical Engineering, 2Center of Imaging, Fuzhou General Hospital of Nanjing Military Command of Chinese PLA, Fuzhou  350025, Fujian Province, China
  • Received:2011-02-12 Revised:2011-03-18 Online:2011-08-06 Published:2011-08-06
  • Contact: Chen Zi-qian, Chief physician, Professor, Doctoral supervisor, Center of Imaging, Fuzhou General Hospital of Nanjing Military Command of Chinese PLA, Fuzhou 350025, Fujian Province, China chenziqianfz@yahoo.con.cn
  • About author:Ni Ping★, Master, Senior engineer, Department of Medical Engineering, Fuzhou General Hospital of Nanjing Military Command of Chinese PLA, Fuzhou 350025, Fujian Province, China nping6@sohu.com
  • Supported by:

    the Social Development Project of Fujian Province, No. 2009J001187*; the Natural Science Foundation of Fujian Province, No. 2009Y0037*; Medical Innovation Project of Nanjing Military Command, No. 09Z029*

PDF

496

被引次数

11

摘要：

背景：细胞的培养时间、数量及纯度是影响干细胞移植的关键因素，优化体外细胞培养模式可以提高移植的成效。
目的：观察不同的首次换液模式、相同的后期换液条件对大鼠骨髓间充质干细胞培养时间及生物学特性的影响。
方法：全骨髓贴壁法培养SD大鼠骨髓间充质干细胞，按首次换液方式分为以下6组：24 h半量换液组、24 h全量换液组、48 h半量换液组、48 h全量换液组、72 h半量换液组、72 h全量换液组，以后每4 h全量换液。记录各组第0~3代培养时间及细胞纯度。
结果与结论：各组培养总时间比较差异有显著性意义(F=46.455，P < 0.05)，48 h全量换液组最短，24 h全量换液组最长，与其他组比较差异均有显著性意义(P < 0.05)。24 h全量换液组各代细胞纯度均最高，72 h半量换液组各代细胞纯度均最低，与其他组比较差异均有显著性意义(P < 0.05)。所获细胞CD29、CD90阳性，CD34、CD45阴性，可以向成骨、成脂诱导分化。结果显示首次换液方式能影响大鼠骨髓间充质干细胞的培养周期及其纯度，48 h半量换液为最佳首次换液方式。

关键词: 首次换液, 贴壁分离, 纯度, 周期, 骨髓间充质干细胞

Abstract:

BACKGROUND: The culture period, quantity and purity are crucial for stem cell transplantation, and it is essential to build an optimal modality of cell culture in vitro. 
OBJECTIVE: To study the influence of different modalities of the first medium exchange on the culture time and biological characteristics of rat bone marrow-derived mesenchymal stem cells.
METHODS: On condition of modality of the first medium exchange, six experimental groups were performed as follows: group A was half exchanged after 24 hours, group B was all exchanged after 24 hours, group C was half exchanged after 48 hours, group D was all exchanged after 48 hours, group E was half exchanged after 72 hours, group F was all exchanged after 72 hours. Then, all groups were all exchanged every 48 hours and were subcultured consecutively. The period and purity of each passage were measured.
RESULTS AND CONCLUSION: The total culture time was significant different (F=46.455, P < 0.001). Group D was the least while group B was the longest, and both of the two groups were significant different compared with others (P < 0.05). Group B earned the highest purity in all passages and group E the lowest, and both of the two groups were significant different compared with others (P < 0.05). Cells of passage 4 showed CD29, CD90 positive, induced to differentiate into osteoblasts and adipocytes. The modality of the first medium exchange had an influence on the culture period and purity of bone marrow-derived mesenchymal stem cells, and the modality of 48-hour half exchange was the best protocol.

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