山羊骨髓间充质干细胞的体外培养与生物学特性

Biologic characteristics of goat bone marrow mesenchymal stem cells cultured in vitro

目的：体外分离培养山羊骨髓问充质干细胞（BMSC），并观察其生物学特性。方法：应用常规密度梯度离心法从山羊骨髓中分离BMSC并传代培养，镜下观察细胞形态及生长特性，绘制细胞生长曲线，测定倍增时间，免疫细胞化学法鉴定其表型。结果：原代培养的BMSC初始呈圆形、梭形及多角形等，细胞呈贴壁生长，48h可见贴壁细胞有伸展现象，细胞渐变为均一梭形，14d时可达90％融合。第1、3和5代BMSC生长曲线呈“s”型，l～2d为潜伏期，2—3d后进入对数增长期，7～8d后进入平台期，平均倍增时间为（23．2±1．9）h。BMSCCD29和CD44呈广泛阳性表达，CD34和CD45呈阴性表达。结论：成功分离培养出山羊BMSC，细胞纯度较高，生长状态良好。

Aim： To isolate and culture goat bone marrow mesenchymal stem cell （BMSC） from bone marrow,and in- vestigate its biologic characteristic. Methods：BMSC was isolated from goat bone marrow by conventional methods. The mor- phology of BMSC was observed with phase contrast microscope, and the growth curve was drawn to determine doubling time. Immunocytoehemical method was used to identify phenotype of BMSC. Results： Primary cultured BMSC was oval, spindle-shaped or polygonal, and adhered to plastic surface within 48 hours, and reached 90% confluence within 14 days. The first, third,and fifth passages of BMSC growth curves showed S-type,and were in latent phase at 1 -2 days,in log phase 2 ~ 3 days later, and came into platform phase at 7 - 8 days. The doubling time was （23.2 ± 1.9） hours. Immunoey- tochemical results showed BMSC was positive for CD29 and CD44,while negative for CD34 and CD45. Conclusion： BMSC has been successfully isolated from goat bone marrow with high purity and activity.