兔骨髓基质干细胞的培养及成骨诱导研究

目的观察体外培养条件下兔骨髓基质干细胞的主要生物学特性及向成骨细胞分化的能力。方法取兔股骨,冲洗骨髓腔,进行细胞的贴壁体外培养,培养至第四代加入含成骨诱导剂的DMEM培养基与空白组为对照组;噻唑蓝（MTT）检测两组骨髓基质干细胞的增殖能力,进行成骨细胞的钙结节茜素红染色与I型胶原的免疫组化染色。结果骨髓基质干细胞7-14 d可见少量的集落形成,24 d时观察见细胞基本铺满瓶底。在矿化液条件下培养1周左右,细胞间出现了致密圆形团,同时细胞的增殖能力较常规培养有所降低;茜素红染色结果显示矿化细胞间出现的致密的、圆形不透光团块呈现片状的棕染,着色区域,I型胶原的免疫组化染色强阳性。结论贴壁筛选法培养骨髓基质干细胞,操作简单,细胞易于成活,不失为一种良好的方法;虽然矿化液可使其向成骨细胞转化,但增殖速度变慢。

The culture of rabbit marrow matrix stem cell and the induction of bone

Objective ： To observe the main biology characteristics of cultural condition of the rabbit marrow matrix stem cell in vitro and the ability to osteoblast. Methods： The rabbit thighbone was selected to flush medullary cavity of bones, for the culture of the cell to paste the wall till the fourth generation, and bone derivative DMEM culture medium was added. The result was compared with that of the control group ; the marrow matrix stem cell multiplication ability in the two groups was examined by MTT, and dyed with the osteoblast the calcium tubercle sodium alizarinsulfonate and I collagen immunity. Results： Marrow matrix stem cell was seen in few colony formations in 7 - 14 d, and the cell was basically full in 24 d. After the culture under the mineralization fluid condition in about 1 wk, the compact circular group appeared in the intercellular space, simultaneously cell multiplication ability reduced compared with the con- ventional culture; The sodium alizarinsulfonate dyeing showed the compact and circular lightproof briquetting brown laminated shape in the mineralizd intercellular spac, I collagen immunity group dyeing was strongly positive in the coloring region. Conclusion ： The method of wall - pasting screening is simple in the cuhutre of marrow matrix stem cell, and the cell is lilkely to survive, which is one good method; Although the mineralization fluid may cause it to transform to the osteoblast, the multiplication speed slows down.