大鼠骨髓基质细胞培养方法的改良

目的　对体外分离培养大鼠骨髓基质细胞 (BMSC)方法进行改良 ,观察其生物学特点。方法　分离大鼠胫骨、股骨 ,以 IMDM培养基冲洗骨髓 ,与培养液混合后直接接种至培养瓶中 ,接种后 7～ 14 d形成单层贴壁的成纤维状细胞。检测传代细胞接种贴壁率、生长曲线、细胞周期 ,电镜观察细胞超微结构。结果　分离培养的原代 BMSC生长良好 ,倍增时间约为 4 2 h。 BMSC传代后 12 h贴壁率达 80 %以上 ,82 %的细胞处于 G0 、G1 期 ,超微结构呈现较早期细胞特点。结论　改良法培养的 BMSC生长稳定 ,传代细胞适应性强。与传统培养方法比 ,该法操作简单、实用性强 ,可以用于 BMSC的体外实验研究

Improvement of the culturing method of rat bone marrow stromal cells in vitro

Objective To develop the method of culturing rat bone marrow stromal cells (BMSC) in vitro, and explore its biological properties. Methods The tibias and femurs of rats were dissencted, the marrow was flushed out with IMDM medium and sent to cultured disk. The cell growth curve, adhensive rate, cell cycle and ultrastructures of passaged cells were tested. Results The adherent, fibroblast-shaped cells approached confluence in single layer 12～16d after plating. The double time of BMSC was about 38h. More than 80% of subcultured BMSC were adhesive in 12h. The cell cycle was analysed and showed that 82% of BMSC was in G 0/G 1 phase. The ultrastructures of BMSC demonstrated the features of infantile cells. Conclusion The subcultured BMSC posses multipotential differentiation and showed table growth, easy survival and rapid proliferation in present culture condition. BMSCs also exhibit infantility in its ultrastructures and cell cycle. The results showed that it may be a useful method for BMSC culture and study of myocardial repairment.