兔骨髓间充质干细胞的体外培养及生物学活性观察

目的建立兔骨髓间充质干细胞(MSCs)体外分离、培养、扩增的方法，并进行生物学活性观察。方法穿刺抽取新西兰大耳白兔骨髓5ml，通过密度梯度离心法获取MSCs，体外贴壁培养、扩增、传代，倒置显微镜下观察原代及各代细胞形态、数量生长情况，描绘生长曲线。第2代细胞用矿化液连续培养后，进行ALP及矿化结节染色。结果体外成功建立兔骨髓间充质干细胞分离扩增传代的方法，能够连续传至8～9代；矿化液培养的细胞ALP及矿化结节染色阳性。结论兔骨髓间充质干细胞能在体外成功培养，具有向成骨细胞诱导分化的特性，可以作为骨组织工程的种子细胞。

Experimental Study on Cultivation of Marrow Mesenchyal Stem Cells in New Zealand White Rabbits in Vitro

Objective To create the model of separation , culture and multiplication of marrow mesenchymal stem cells from New Zealand white rabbits, and observe their biological features. Methods 5 ml bone marrow was drawn by puncture from New Zealand white rabbits. Morrow mesenchymal cells were harvested through gradient centrifugation, then were cultivated, multiplied and expanded. Primary cells and sub-cultured cells were estimated through observing their morphology and number, drawing growth curve. After cultivated continuously in mineralizational medium for 3 weeks, the cells of the fourth passage were stained by calcifying nodule Vonkossa and alkaline phos-phatase(ALP) Calcium-Cobalt methods. Results The model of culture and expansion of BMSCs in vitro were established. Bone marrow mesenchymal stem cells were induced to osteoblasts successfully in vitro. Conclusion Marrow mesenchymal stem cells can be cultivated successfully in vitro, and possess the feature to be induced to osteoblasts and can serve as excellent seeding-cells for bone tissue engineering .