中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (6): 957-961.doi: 10.3969/j.issn.1673-8225.2010.06.002

• 干细胞培养与分化 • 上一篇    下一篇

间充质干细胞在低氧环境下的增殖、代谢与成骨分化：胎盘羊膜及骨髓来源间充质干细胞的对比

陈 亭1，周 燕1，张治萍2，谭文松1   

  1. 1华东理工大学动物细胞与组织工程研究室，上海市  200237；
    2上海市浦东新区妇幼保健院，上海市  201200
  • 出版日期:2010-02-05 发布日期:2010-02-05
  • 通讯作者: 周 燕，硕士，副教授，华东理工大学动物细胞与组织工程研究室，上海市 200237 zhouyan@ecust.edu.cn
  • 作者简介:陈 亭，男，1982年生，江苏省徐州市人，汉族，华东理工大学在读博士，主要从事干细胞扩增与分化的研究。 shengwu01@163.com
  • 基金资助:

    微组织体外构建技术及其过程分析，项目批准号：20776044，国家自然科学基金项目。

Proliferation, metabolism, and osteogenic differentiation of mesenchymal stem cells under hypoxia: Cell sources of placenta amniotic versus bone marrow

Chen Ting1, Zhou Yan1, Zhang Zhi-ping2, Tan Wen-song1   

  1. 1Laboratory of Animal Cells and Tissue Engineering, East China University of Science and Technology, Shanghai   200237, China;
    2Maternal and Child Health Care Hospital in Shanghai Pudong New Area, Shanghai   201200, China
  • Online:2010-02-05 Published:2010-02-05
  • Contact: Zhou Yan, Master, Associate professor, Laboratory of Animal Cells and Tissue Engineering, East China University of Science and Technology, Shanghai 200237, China zhouyan@ecust.edu.cn
  • About author:Chen Ting, Studying for doctorate, Laboratory of Animal Cells and Tissue Engineering, East China University of Science and Technology, Shanghai 200237, China shengwu01@163.com
  • Supported by:

    the National Natural Science Foundation of China, No. 20776044*

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摘要：

背景：目前关于低氧对间充质干细胞成骨分化影响的报道结果有差异，这些差异可能是采用氧体积分数不同引起的，也有可能是间充质干细胞来源不同引起的。

目的：比较胎盘羊膜间充质干细胞和骨髓间充质干细胞在低氧下的生物性状差异。

方法：两步酶消化法与全骨髓贴壁法分别获得人胎盘羊膜间充质干细胞和兔骨髓间充质干细胞，体外传代培养。分别取相同代次的人胎盘羊膜间充质干细胞和骨髓间充质干细胞，细胞以2×104 cells/孔接种于12孔板，加入含体积分数为10%胎牛血清的α-MEM培养基，分别在体积分数为5%O2或体积分数为20%O2条件下培养生长12 d。成骨诱导时，细胞以1×105 cells/孔接种于12孔板，加入成骨诱导培养基，分别在体积分数为5%O2或体积分数为20%O2条件下成骨诱导14 d。观察两种间充质干细胞在不同氧浓度下的生长情况及在不同氧浓度下葡萄糖的消耗与乳酸生成及成骨分化情况。

结果与结论：与常氧组相比，低氧促进人胎盘羊膜间充质干细胞和骨髓间充质干细胞的增殖和成骨分化。但与骨髓间充质干细胞相比，低氧能更好地促进人胎盘羊膜间充质干细胞的增殖；低氧下人胎盘羊膜间充质干细胞消耗较少的葡萄糖，产生较少的乳酸。在成骨诱导方面，低氧显著促进骨髓间充质干细胞的碱性磷酸酶表达，且远高于人胎盘羊膜间充质干细胞，但是两者的单位细胞钙沉积量无明显差异。

关键词: 低氧, 胎盘羊膜间充质干细胞, 骨髓间充质干细胞, 增殖, 代谢, 成骨分化

Abstract:

BACKGROUND: Effect of hypoxia on osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) has been differently reported. Those differences might cause by varying volume fraction of oxygen and varying source of BMMSCs.

OBJECTIVE: To compare the biological differences between placenta and BMMSCs under hypoxia.

METHODS: Human placenta amniotic mesenchymal stem cells (hAMSCs) and rabbit BMMSCs were isolated by two step proteinases and whole bone marrow adhesion, respectively. hAMSCs and BMMSCs at the same passage were seeded in 12-well plates at an initial cell density of 2 × 104 cells per well with α-MEM containing 10% FBS. Then, the cells were cultured under 5% O2 or 20% O2 for 12 days. hAMSCs and BMMSCs at the same passage were seeded in 12-well plates at an initial cell density of 1 × 105 cells per well with osteogenic medium. Then, the cells were cultured under 5% O2 or 20% O2 for 14 days. Cell growth curve, the specific glucose consumption rates and specific lactate production rates, and osteogenic differentiation were detected.

RESULTS AND CONCLUSION: Compared to normal oxygen, hypoxia promoted the proliferation and osteogenic differentiation of MSCs. When compared to BMMSCs, statistically significant enhancement of the growth of hAMSCs by hypoxia was observed. hAMSCs cultured under hypoxia exhibited lower glucose consumption and lactate production in contrast with BMMSCs. Furthermore, comparison between hAMSCs and BMMSCs showed that the alkaline phosphatase expression of BMMSCs was significantly enhanced by hypoxia and was markedly higher compared with hAMSCs. The amount of calcium deposition was also enhanced by hypoxia, but there were no statistically significant differences between hAMSCs and BMMSCs.

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