与软骨细胞共培养和条件培养液诱导骨髓间充质干细胞向软骨细胞分化的比较

背景：骨髓间充质干细胞体外转化很大程度上依赖于合适的培养条件。目的：比较与软骨细胞共培养和条件培养液2种不同的诱导方案诱导骨髓间充质干细胞向软骨细胞分化的特点。方法：分离培养大鼠骨髓间充质干细胞和耳软骨细胞,采用骨髓间充质干细胞与软骨细胞共培养及条件培养液诱导成软骨的方法,诱导骨髓间充质干细胞向软骨细胞分化。以MTT法及流式细胞仪检测细胞活性及周期,糖胺多糖、甲苯胺蓝以及免疫组化染色检测细胞生物学特性,以RT-PCR法检测诱导后的软骨细胞Ⅱ型胶原RNA表达情况。结果与结论：采用共培养方式诱导的软骨细胞,其生物学特性与采用条件培养液诱导的软骨细胞相比,前者优于后者,如分泌糖胺多糖的能力以及基质分泌量均较高。提示共培养方式诱导的软骨细胞更接近正常软骨细胞,更有利于作为组织工程软骨的种子细胞。

A comparative study on bone marrow mesenchymal stem cells differentiated into chondrocytes induced by co-culture with chondrocytes and conditioned medium

BACKGROUND： In vitro differentiation of bone marrow mesenchymal stem cells （BMSCs） is dependent on appropriate culture conditions to a great extent.OBJECTIVE： To study the biological characteristics of chondrocytes differentiated from BMSCs co-cultured with chondrocytes versus cultured in conditioned medium.METHODS： Rat BMSCs and chondrocytes were isolated and cultured in neural induction.Cellular non-contact co-culture induced method and conditioned medium induced method were taken to reduce BMSCs differentiating into chondrocytes.And the biological characteristics of the cells were evaluated by MTT,mitotic cycle,GAG secretion,RT-PCR,cell-staining methods,immunocytochemistry.RESULTS AND CONCLUSION： The biological characters of co-culture induced cells were seemed to be better than those of conditioned medium induced cells,which point out that BMSCs co-cultured with chondrocytes are appropriate as seed cells in cartilage tissue engineering.