Isolation and culture of human bone marrow mesenchymal stem cells without lysing the erythrocytes / 中国综合临床

ABSTRACT

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-NH4Cl 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 l07 cells were compared between the two groups.Results The times for P1,P2 and P3 passage was ( 9.3 ± 4.9 ) days vs.( 7.2 ± 1.0 ) 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 =0.06,t =0.80,P ＞ O.05 ).The time for expansion to 107 cells in two groups was not significantly different ( t =0.80,P ＞0.05).Conclusion Tris-NH4Cl 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.