摘要
目的建立大鼠骨髓间质干细胞(MSCs)最佳的体外分离培养方法,并研究其基本的生物学特性。方法采用密度梯度离心和贴壁培养两种方法分离大鼠的MSCs,进行形态学观察及增殖生长方面的测定;免疫细胞化学染色鉴定表面标志。结果大鼠MSCs体外培养呈梭形;细胞生长曲线为“S”型;密度梯度离心法原代培养15d达到80%～90%的融合,而改良的直接贴壁法只需要5d;细胞周期显示G0/G1期88.29%;免疫细胞化学染色显示CD44表达阳性,CD34、CD45为阴性。结论两种方法均可获得骨髓间质干细胞,但贴壁培养法操作更简便、快速,具有对细胞活性影响较小,更利于其贴壁和增殖的优点。MSCs体外培养条件下生长性状稳定,适于做组织工程的种子细胞。
Objective To seek the best way of isolating and culturing rat bone marrow mesenchymal stem cells (MSCs) and to investigate the biological characteristics of rat bone marrow MSCs. Methods MSCs were obtained with density gradient centrifugation and direct adherence separately, and their living characteristics were observed. Immunocytochemical technique was performed to examine the specific antigens on the MSCs. Results The cultured cells were spindle-shaped. The growth curve liked “S” shape. Primary cultured cells could attained 80%～90% integral for 15 days through density gradient centrifugation,while ameliorated direct adherence need only 5 days.The cell cycle analysis showed that 88.29% of MSCs was in G_0/G_1 phase. The cultured cells express CD44, but did not express CD34、CD45. Conclusion Both the above-mentioned way can obtain MSCs, but the adherence was more easily operated,it had little influence to cells activity and was better for adherence and proliferation. So, MSCs can be used as the seed cells for tissue engineering.
出处
《山西医科大学学报》
CAS
2005年第2期158-161,共4页
Journal of Shanxi Medical University
关键词
骨髓
间质干细胞
细胞培养
bone marrow
mesenchymal stem cells
cell culture
