早期贴壁分离并机械刮除纯化法建立兔骨髓间充质干细胞体外培养体系

①目的探讨早期（60-80分钟）贴壁分离法建立兔骨髓间充质干细胞体外培养体系，对细胞贴壁分离的时机以及传代后的形态变化的影响。②方法采用早期贴壁分离方法及机械刮除纯化法纯化问充质细胞，并用钙钴法染色检测细胞的ALP表达情况、使用成骨诱导荆诱导间充质细胞。用茜素红染色观察矿化结节的形成。③结果第一代细胞碱性磷酸酶染色呈阳性；经条件培养基定向培养5天的细胞呈集落生长，并形成钙结节，茜素红染色阳性。④结论采用早期贴壁分离法可以短期内得到大量较纯的免间充质干细胞。在诱导剂作用下可成功分化成为成骨细胞，是培养骨组织工程用途的种子细胞的理想方法。

In vitro culture MSCs by isolating the cells of anchoring at earlier period and physically curetlementing

Objective To culture the Mesenchymal stem cells(MSCs) of rabbits in vitro,studying the change in morphology of passage cells and the occasion of isolating the anchoring cells.Method The bone marrow mesenchymal stem cells of New Zealand rabbits were cultured.The MSCs were physically curetlemented and purified by the way of isolating the cells of anchoring at early phage(one hour).The expression of ALP in cells were messured.The MSCs were induced by osteogenisis opponents.The formation of the mineralization nod was observed by chinalizarin staining.Results MSCs showed the positive expression of alkaline phosphates after being cultured in 2 weeks;The localized regions of mineralization were found at about 5 days with conditioned culture medium.Conclusions The comparatively purified rabbits MSCs are obtained massly in short period by isolating the anchoring cells of one hour.The MSCs can be induced into osteoblast cells successfully by inducing components.The method is a optimal way in culturing seed cells in the bone tissue engineering.