人骨髓间充质干细胞体外分离无需红细胞裂解  

Isolation and culture of human bone marrow mesenchymal stem cells without lysing the erythrocytes

作　　者：陈国军[1] 王亚莉[1] 方凤[1] 刘厚奇[2] 

机构地区：[1]武警浙江省总队医院细胞治疗中心,嘉兴314000 [2]第二军医大学发育生物学研究所

出　　处：《中国综合临床》2012年第4期344-346,共3页Clinical Medicine of China

基　　金：国家重点基础研究发展计划项目（201lCB965101）

摘　　要：目的观察红细胞裂解液对人骨髓间充质干细胞（hMSCs）体外分离、培养及增殖特性的影响。方法将22份骨髓随机分为两组：（1）碎红组11份，即骨髓样本在处理过程中给予Tris-NH。C1红细胞裂解剂；（2）未碎红组11份，即骨髓样本处理过程中未加红细胞裂解剂。观察两组细胞生长情况，第1、2、3代细胞达到传代的时间和细胞扩增达到107数量的时间。结果（1）碎红组与未碎红组在P1传代时间分别为（9．3±4．9）d和（7．2-4-1．0）d、P2传代时间（14．4±4．7）d和（14．5±3．5）d、P3传代时间（18．5±5．0）d和（20．1±4．4）d，两组在传代时间上差异均无统计学意义（f值分别为1．39、0．06、0．80，P均〉0．05）；（2）碎红组与未碎红组在细胞培养达到10’数量的时间差异无统计学意义[（19．5±7．9）、（22．1±7．3）d，t=0．80，P〉0．05]。结论添加与不添加Tris-NH4C1红细胞裂解剂对hMSCs的体外分离、培养和细胞增殖速度方面分析均无影响。故hMSCs体外分离培养可省略红细胞裂解步骤。Objective To observe the effects of red blood cell lysis buffer on the isolation and culture of human bone marrow mesenchymal stem cells （hMSCs） in vitro. Methods Twenty-two bone marrow samples were randomly divided into 2 groups, including 11 samples of crushed red blood cell prepared with Tris-NH4 C1 red blood cell lysis agent and 11 samples prepared without red blood cell lysis agent. The intervals from primary generation to 1,2 and 3 passages and the time of expansion to 107 cells were compared between the two groups. Results ThetimesforP1,P2 and P3 passage was （9.3 ±4.9）days vs. （7.2±l.0）days,（14.4± 4. 7） days vs. （ 14. 5 ± 3.5 ） days, and （ 18.5 ± 5.0） days vs, （ 20. 1 ± 4.4 ） days, respectively, in crushed red blood cell group and non-crushed red blood cell group. The differences were not significant （ t = 1.39, t = O. 06, t =0. 80,P 〉0. 05）. The time for expansion to 107 cells in two groups was not significantly different （t =0. 80, P 〉 0. 05）. Conclusion Tris-NH6 C1 agent in red blood cell lysis has no significant effects on hMSCs isolation, culture and cell proliferation, which indicates erythrocyte lysing may be not an independent step for hMSCs isolation and expansion in vitro.

关 键 词：骨髓间充质干细胞 红细胞裂解液 分离 培养 增殖 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]