儿童骨髓基质细胞体外培养

目的 探讨儿童骨髓基质细胞体外培养条件，为儿童骨组织工程研究选择种子细胞。方法 将儿童髂骨内骨髓分别采用密度梯度离心法和全骨髓法培养，待细胞长满培养瓶后传代，选取正常生长骨髓基质细胞第3代绘制生长曲线并测定细胞内骨钙素(OCN)及培养液中碱性磷酸酶(ALP)含量。培养期间用倒置相差显微镜观察细胞形态。结果 原代儿童骨髓基质细胞呈长梭形、三角形或多角形，存在有集落样生长特性，传代后细胞形态无明显变化，且生长、增殖稳定，细胞内OCN及培养液中ALP含量在1周左右达到高峰。结论 采用密度梯度离心法和全骨髓法获得儿童骨髓基质细胞，经培养、传代后，可以得到大量稳定增殖的骨组织工程种子细胞。

Culture of children marrow stromal cells in vitro

Objective To explore the culture of children marrow stromal cells (MSCs) in vitro for the bone tissue researching.Methods We got the iliac bone marrow and cultured the cells by density gradient centrifugation method and whole bone marrow method.After the transfer of culture,we drew the cell growth curve of third generation and determined the OCN in the MSCs and ALP in the culture medium,and observed the cell form during the culture.Results The MSCs appeared long fusiform,triangle and polygonal and also presented the characteristic of colony growth.It did not change after transfer of culture,and the growth and proliferation was stable.The OCN in the MSCs and ALP in the culture medium reached peak value at about one week after culturing.Conclusion We can get lots of stable MSCs by density gradient centrifugation method and whole bone marrow method for bone tissue engineering.