不同冻存方法对大鼠脂肪来源干细胞生物学特性的影响

Effects of Different Cryopreservation Methods on Biological Characteristics of Adipose Stem Cells in Rats

目的:研究不同冻存方法对脂肪来源干细胞(Adipose-derivedstemcells,ADSCs)生物学特性的影响。方法:体外分离培养SD大鼠腹股沟及附睾周围的脂肪组织,取第三代ADSCs冻存于液氮。实验分为三组:对照组A组为非冻存细胞,B组使用无血清非程序冻存液冻存细胞,C组使用传统冻存液(DMEM∶FBS∶DMSO=5∶4∶1)冻存细胞。12个月后复苏ADSCs,通过对比ADSCs的表面抗原、细胞凋亡、增殖及成骨分化情况,分析各组ADSCs生物学性能的差异。结果:各组细胞高表达CD29、CD90;A、B两组细胞的凋亡率显著低于C组(P<0.05);B、C两组的增殖情况和A组相比,差异无统计学意义(P>0.05),B组细胞的成骨分化能力要优于C组(P<0.05)。结论:无血清非程序冻存液冻存ADSCs的效果要优于传统冻存液。

Objective To study the efiects of difierent cryopreservation methods on the biological characteristics of ADSCs.Methods Tissue from groin and epididymis of SD rats was isolated and cultured invitro,the third passage cells were frozen in liquid nitrogen.The experiment was divided into three groups:control group A was non-cryopreservation cells,In group B,cells were cryopreserved with serum-free non-programmed cryopreservation method,The cells in group C were cryopreserved with conventional cryopreservation method(DMEM∶FBS∶DMSO=5∶4∶1).ADSCs were resuscitated after 12 months,Through contrast the surface antigen of ADSCs,cell apoptosis,cell proliferation assay and osteogenic difierentiation results.To analyse the difierence of ADSCs in each group.Results CD29 and CD90 were highly expressed in each group.The apoptosis rate of cells in group A and B was significantly lower than that in group C(P<0.05).There was no significant difierence in proliferation between group B and group C(P>0.05).The osteogenic difierentiation ability of group B was better than that of group C(P<0.05).Conclusion The efiect of serum-free non-programmed cryopreservation method on ADSCs was better than that of conventional cryopreservation method.