贾杰;杨述华;张宇坤;孙志博;

• 1:华中科技大学同济医学院附属协和医院骨科

摘要(Abstract)：

[目的]探究以人骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hMSCs)为种子细胞体外构建"自组装"工程化软骨对应的合理细胞接种密度。[方法]体外分离与培养hMSCs。用含100 ng/ml生长分化因子5(growth differentiation factor 5,GDF-5)的软骨诱导液(chondrogenic medium,CM)定向诱导培养第3代hM-SCs,诱导3周后重悬细胞,分别以A组2.5×106/ml,B组5×106/ml,C组1×107/ml,D组2×107/ml四种细胞密度接种于2%琼脂糖包被的24孔板,每组设5个复孔。自组装培养3周后,对标本进行大体观察、组织学及免疫组化检测并进行生化分析,比较不同组标本的软骨生物学特性的差异。[结果]"自组装"培养3周后,各组都形成了软骨样组织团块,团块直径与湿重随细胞接种密度而增加。Bern评分结果显示C、D两组明显高于A、B两组(P<0.05),且C组与D组间差异无统计学意义,Ⅱ型胶原免疫组化染色检测到C组与D组细胞外基质内有较强的阳性信号弥漫分布,蛋白多糖(GAG)含量C、D两组亦明显高于A、B两组(P<0.05),且C组与D组间差异无统计学意义。[结论]在一定细胞接种密度范围内,"自组装"工程化软骨的生物学特性呈密度依赖性增加。以1×107/ml接种时,可以获得具有良好生物学特性的"自组装"工程化软骨。

关键词(KeyWords)： 成人骨髓间充质干细胞;生长分化因子5;“自组装”培养;软骨分化

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(编号:30800654)

作者(Author): 贾杰;杨述华;张宇坤;孙志博;

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