摘要
目的比较两种不同方法分离兔骨髓间充质干细胞在体外培养环境下的生长增殖及表型特点,为软骨组织工程提供种子细胞。方法行全骨髓培养法和梯度密度离心培养法分离骨髓间充质干细胞,用倒置显微镜、透射电镜观察细胞生长状况,3H标记的胸腺嘧啶脱氧核苷(3H-TdR)掺入实验检测细胞的增殖情况,对骨髓间充质干细胞进行表型鉴定。结果全骨髓培养法的骨髓间充质干细胞,原代培养周期为14天,原代及传代细胞一致表达CD44,细胞3H-TdR掺入实验每分钟脉冲数为15250cpm;梯度密度离心法的骨髓间充质干细胞,原代培养周期为28天,原代及传代细胞表达CD44,细胞3H-TdR掺入实验每分钟脉冲数为13570cpm。结论用全骨髓培养法培养骨髓间充质干细胞不失为一种很好的方法。
Objectives To compare the proliferation and surface antigen ofmesenchymal stem cells isolated by two different method from rabbit bone marrow. And the stem cell will be used as potential seeding ceils in cartilage tissue engineering. Methods MSCs were isolated from the Percoll fraction ofmononuclear cells and whole bone marrow cells.cells were examed grossly, histologically and electron microscoply.The proliferation of the cells was examined by ^3H -TdR incorporation. Expression of cell surface molecules on BMSCs were determined by flow cytometry. Result The primary culture period of the whole bone marrow and adhere cultured stem cell is about 14d. CD44 are expressed on almost all the primary and the following passage cell. The sequences of the incorporation of ^3H -TdR were 15250cpm; The primary culture period of the BMSCs isolated from the Percoll fraction of mononuelear cells is about 28d. CD44 are expressed on part of the primary and the following passage cells. The sequences of the incorporation of ^3H -TdR were 13570cpm. Con- clusions The combination of whole bone marrow and adhere culture is an effective method to isolate BMSCs from bone marrow aspirate.
出处
《生物骨科材料与临床研究》
CAS
2008年第5期1-3,共3页
Orthopaedic Biomechanics Materials and Clinical Study
基金
山西医科大学第二医院青年基金项目(编号:20070103)
关键词
骨髓间充质干细胞
细胞培养
Bone marrow mesenchymal stem cells
Cell culture
