中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (32): 5926-5930.doi: 10.3969/j.issn.2095-4344.2012.32.007

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

郑 亮，李萍华，刘钰瑜，崔 燎   

  1. 广东医学院药理学教研室，广东省湛江市 524023
  • 收稿日期:2011-12-17 修回日期:2012-03-28 出版日期:2012-08-05 发布日期:2012-08-05
  • 通讯作者: 刘钰瑜，硕士，副教授，硕士生导师，广东医学院药理学教研室，广东省湛江市 524023 liuyuyu77@163.com
  • 作者简介:郑亮，女，1985年生，广东省湛江市人，汉族，2009年广东医学院毕业，主要从事抗骨质疏松的研究。 willspare@163.com

Induced differentiation of bone marrow mesenchymal stem cells into adipocytes in vitro

Zheng Liang, Li Ping-hua, Liu Yu-yu, Cui Liao   

  1. Department of Pharmacology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China
  • Received:2011-12-17 Revised:2012-03-28 Online:2012-08-05 Published:2012-08-05
  • Contact: Liu Yu-yu, Master, Associate professor, Master’s supervisor, Department of Pharmacology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China liuyuyu77@163.com
  • About author:Zheng Liang, Department of Pharmacology, Guangdong Medical College, Zhanjiang 524023, Guangdong Province, China willspare@163.com

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摘要：

背景：骨髓间充质干细胞可向成骨细胞、脂肪细胞等多个方向分化，建立稳定可靠的诱导骨髓间充质干细胞成脂分化的方法可为该细胞用于骨质疏松症等骨骼系统相关疾病的防治研究提供保证。
目的：探讨骨髓间充质干细胞在体外定向分化为脂肪细胞的适宜诱导条件。
方法：无菌条件下分离大鼠股骨，密度梯度离心法与贴壁培养法相结合分离、纯化骨髓间充质干细胞并在体外进行培养。第3代骨髓间充质干细胞完全汇合后，利用含有3-异丁基-1-甲基黄嘌呤、吲哚美辛、地塞米松、胰岛素及胎牛血清的DMEM低糖培养液定向诱导骨髓间充质干细胞分化为脂肪细胞。
结果与结论：①倒置显微镜下观察，传3代后可获得均一性较高的骨髓间充质干细胞，经过诱导可定向分化成脂肪细胞。②在3-异丁基-1-甲基黄嘌呤、吲哚美辛、地塞米松诱导的条件下，采取间断诱导方法，骨髓间充质干细胞在体外可定向分化为脂肪细胞。③实验中用5×10-4 mol/L IBMX，2×10-4 mol/L吲哚美辛，10-4 mol/L地塞米松诱导间充质干细胞的成脂分化效果较好。提示骨髓间充质干细胞经含0.1 mmol/L 3-异丁基-1-甲基黄嘌呤、10 mg/L胰岛素、 0.1 mmol/L吲哚美辛、1 μmol/L地塞米松、体积分数为10%胎牛血清的DMEM分化诱导液定向诱导可获得较好的成脂分化效果。

关键词: 骨髓间充质干细胞, 脂肪细胞, 分化, 骨质疏松, 正交设计, 干细胞

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are able to differentiate into different cell lineages such as adipocytes and osteoblastes. Establishing a stable and reliable induction method to induce BMSCs to differentiate into adipocytes is of great help in studying prevention and treatment of osteoporosis and skeletal-related diseases.
OBJECTIVE: To investigate the optimal conditions for induced differentiation of BMSCs into adipocytes.
METHODS: Under aseptic condition, rat femur was harvested. BMSCs were isolated and purified by density-gradient centrifugation and adherent culture in vitro. After complete confluency, passage 3 BMSCs were induced to differentiate into adipocytes using DMEM containing 3-isobutyl-1-methylxanthine, indomethacin, dexamethasone, insulin and fetal bovine serum.
RESULTS AND CONCLUSION: Under the inverted microscope, highly homogenous BMSCs could be harvested after three passages, and these cells could be induced to differentiate into adipocytes. BMSCs could be committedly to differentiate into adipocytes in vitro after induced by 3-isobutyl-1-methylxanthine, indomethacin, dexamethasone. 5×10-4 mol/L 3-isobutyl-1-methylxanthine, 2×10-4 mol/L indomethacin and 10-4 mol/L dexamethasone exhibited better effects on induced differentiation of BMSCs into adipocytes. The best condition for inducing BMSCs to differentiate into adipocytes in vitro was DMEM supplemented with 0.1 mmoL/L 3-isobutyl-1-methylxanthine, 10 mg/L insulin, 0.1 mmoL/L indomethacin, 1 μmoL/L dexamethasone, and 10% fetal bovine serum.

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