中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (27): 5037-5040.doi: 10.3969/j.issn.1673-8225.2010.27.022

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

骨髓间充质干细胞和关节软骨细胞共培养与转化生长因子β1诱导的比较

陈宗雄，刘晓强，王万宗   

  1. 解放军南京军区福州总医院骨一科，福建省福州市  350025
  • 出版日期:2010-07-02 发布日期:2010-07-02
  • 通讯作者: 王万宗，博士，主治医师，解放军南京军区福州总医院骨一科，福建省福州市 350025 jfj999999@live.cn
  • 作者简介:陈宗雄☆，男，1975年生，福建省莆田市人，汉族， 2000年解放军第二军医大学毕业，博士，副主任医师，主要从事脊柱关节疾病研究。 Lxq999999@live.cn

Co-culture bio-induction of bone marrow mesenchymal stem cells and chondrocytes versus transforming growth factor beta 1 chemical induction

Chen Zong-xiong, Liu Xiao-qiang, Wang Wan-zong   

  1. First Department of Orthopaedics, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou  350025, Fujian Province, China
  • Online:2010-07-02 Published:2010-07-02
  • Contact: Wang Wan-zong, Doctor, Attending physician, First Department of Orthopaedics, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou 350025, Fujian Province, China jfj999999@live.cn
  • About author:Chen Zong-xiong☆, Doctor, Associate chief physician, First Department of Orthopaedics, Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA, Fuzhou 350025, Fujian Province, China Lxq999999@live.cn

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摘要：

背景：应用生长因子可诱导骨髓间充质干细胞向软骨样细胞分化，但诱导后的细胞在生物体内很难形成成熟的软骨细胞且仍具有分泌软骨基质及抗压，抗摩擦的能力。
目的：对比分析骨髓间充质干细胞与关节软骨细胞共培养诱导、转化生长因子β1诱导骨髓间充质干细胞的效果。
方法：提取SD大鼠关节软骨细胞与第3代骨髓间充质干细胞，按1∶2，1∶1，2∶1浓度比种植于Transwell共培养系统中。同时设置转化生长因子β1 诱导组为对照。相差显微镜下观察细胞的增殖和基质合成情况，并诱导结果行MTT比色法检查、氨基聚糖水平检测及Western Blot检测Ⅱ型胶原基因的表达情况。
结果与结论：关节软骨细胞与骨髓间充质干细胞1∶2比例诱导的结果与10 µg/L 转化生长因子β1诱导结果相当，但随着关节软骨细胞在诱导中体系中比例的增加，诱导结果明显优于转化生长因子β1诱导，当诱导细胞达到一定比例时，诱导结果不会随之变化。说明软骨细胞与骨髓间充质干细胞共培养可以诱导骨髓间充质干细胞向软骨细胞转化，在开始时，诱导结果与软骨细胞所占的比例成正相关，当软骨细胞达到一定比例时， 骨髓间充质干细胞的诱导结果并未发生明显变化，提示骨髓间充质干细胞对软骨细胞的诱导存在饱和现象。

关键词: 骨髓间充质干细胞, 关节, 软骨细胞, Transwell共培养, 软骨样诱导, 转化生长因子&beta, 1

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can differentiate into chondrocytes-like cells by factor induction. However, induced cells difficultly form mature chondrocytes in an organism, and still have the ability of secreting cartilage matrix, anti-pressure and anti-friction.
OBJECTIVE: To compare and analyze the effect of the induction of BMSCs by co-culture BMSCs and chondrocytes induction and transforming growth factor (TGF)-β1 induction.
METHODS: Chondrocytes and the third passage BMSCs were harvested from Sprague Dawley rats, and incubated in Transwell co-culture system at 1: 2, 1: 1 and 2: 1. Simultaneously, TGF-β1 induction group served as a control. Cell proliferation and matrix synthesis would be observed under a phase contrast microscopy. MTT assay, glucose amino glycan (GAG) content detection and Western Blot assay were adopted to check the expression of type Ⅱ collagen gene.
RESULTS AND CONCLUSION: The result of BMSCs and chondrocytes (1: 2) induction was quite the same as that of 10 μg/L TGF-β1 induction, but as the increasing percentage of chondrocytes in induction, the inductive result obviously has an advantage over TGF-β1 induction. When induced cells come to some rate, the result would not change, which proved that co-culture BMSCs and chondrocytes can induce BMSC transformation into chondrocytes. The induction result was positively related to the percentage of chondrocytes at the beginning; when chondrocytes come to some rate, the result of BMSCs induction does not significantly change. Above-mentioned results have shown that BMSC induction into chondrocytes exists saturation.

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