中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (6): 989-993.doi: 10.3969/j.issn.1673-8225.2012.06.009

• 脐带脐血干细胞 umbilical cord blood stem cells • 上一篇    下一篇

梯度血清递减体外培养人脐带间充质干细胞

李  力，庞  妍，张  航，邝丽萍，肖芷芳，王耀春，肖  扬   

  1. 解放军广州军区广州总医院，广东省广州市 510010
  • 收稿日期:2011-07-17 修回日期:2011-08-14 出版日期:2012-02-05 发布日期:2012-02-05
  • 作者简介:李力，女，1969年生，湖北省武汉市人，汉族，1992年解放军第三军医大学毕业，副主任技师，主要从事间充质干细胞及肿瘤免疫细胞的研究。Lily17155@yahoo.com.cn

R394.2Human umbilical cord mesenchymal stem cells cultured in serum-decremental medium in vitro

Li Li, Pang Yan, Zhang Hang, Kuang Li-ping, Xiao Zhi-fang, Wang Yao-chun, Xiao Yang   

  1. General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou  510010, Guangdong Province, China
  • Received:2011-07-17 Revised:2011-08-14 Online:2012-02-05 Published:2012-02-05
  • About author:Li Li, Associate chief technician, General Hospital of Guangzhou Military Command of Chinese PLA, Guangzhou 510010, Guangdong Province, China Lily17155@yahoo.com.cn

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286

被引次数

1

摘要：

背景：人脐带间充质干细胞的扩增与培养条件密切相关，而含体积分数10%~20%胎牛血清的培养基可促进细胞生长。
目的：建立梯度血清递减扩增培养脐带间充质干细胞的培养技术。
方法：采用胶原酶Ⅱ消化分离获得脐带间充质干细胞悬液，并通过贴壁培养进行纯化，细胞贴壁后采用梯度血清递减方法，即第1代80%含血清培养基，20%无血清培养基；第2代50%含血清培养基，50%无血清培养基；第3代20%含血清培养基，80%无血清培养基；第4代100%无血清培养基。另一种传代中始终采用含体积分数10%胎牛血清的α-MEM完全培养液。用流式细胞仪检测细胞的表面标记，进行成骨诱导试验，同时与经典的含10%胎牛血清的α-MEM培养体系进行比较。
结果与结论：梯度血清递减培养法与经典α-MEM培养体系所获得的细胞在扩增能力、细胞形态、免疫表型等方面相似。细胞在两种培养体系中均能保持良好的分化潜能。但梯度血清浓度法培养间充质干细胞可使用更少量胎牛血清，提高临床应用安全性。

关键词: 胎牛血清, 脐带, 间充质干细胞, 培养, 扩增

Abstract:

BACKGROUND: The proliferation of human umbilical cord mesenchymal stem cells (hUCMSCs) strongly depends on the culture conditions. Medium containing 10-20% fetal bovine serum (FBS) can promote cell growth.
OBJECTIVE: To build the serum-decremental technology for UCMSCs culture.
METHODS: hUCMSCs were separated using collagenase Ⅱ digestion method and purified by adherent culture. The cells were cultured and expanded by the serum-decremental method: media 20% serum-free medium for the primary culture, 50% serum-free medium for the second passage, 80% serum-free medium for the third passage, and 100% serum-free medium for the fourth passage. Cells were cultured in α-MEM medium containing 10% FBS as control. Cell surface markers were tested by flow cytometry. Osteogenic induction test was performed.
RESULTS AND CONCLUSION: Cells cultured in the both media were similar in proliferative ability, morphology, and immunophenotype. Their differentiative potential kept well in both media. However, less FBS was used in serum-decremental medium. Serum-decremental medium improves the safety of hUCMSCs culture in clinical practice.

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