克隆化兔骨髓基质干细胞系的建立及其生物学特性研究

目的 建立克隆化兔骨髓基质干细胞（MSC）系，并对其生物学特性进行研究，为组织缺损修复寻找理想的种子细胞提供新的途径。方法 应用密度梯度离心和贴壁分离培养方法，从兔骨髓中获得原代骨髓MSCs，再应用克隆化分离培养技术，建立克隆化扩增的骨髓MSC系。体外传代培养、低温冻存复苏、形态学观察、生长曲线测定、染色体分析和分化潜能鉴定。结果 建立了一株克隆化兔骨髓MSC系。该细胞系在含10％胎牛血清的LG-DMEM中连续传代至第32代，仍保持旺盛的增殖能力，P4和P32代细胞染色体众数保持稳定的二倍体核型，成骨细胞诱导试验阳性。结论 获得一株具有旺盛的增殖能力和分化潜能，遗传性能稳定的克隆化兔骨髓MSC系。

Establishment and Characterization of Clonal Mesenchymal Stem Cell Line from Bone Marrow of Rabbit

Objective To establish cloned bone marrow mesenchymal stem cell line, and analysis the characteristics. Methods MSC was isolated from rabbit bone marrow by density gradient centrifugation and their preferential adherence to the culture dish. The cells of the primary culture were cloned and a cell line was established. The cell line was cultured in vitro by consecutive passage or recovering from the frozen ones.The characteristics such as morphology, growth curve, chromosome and differentiation potential,were analyzed. Results After 32 passages, the cell line was still able to proliferate vigorously, the original karyotype was kept, and the osteoblast induction assay was positive.Conclusion The established MSC line had high proliferation and differentiation potentials and was genetically stable.