不同培养方法对小鼠脂肪干细胞成骨分化的影响

Influence of different culture methods on osteogenic differentiation of adipose-derived stem cells

目的比较组织块贴壁法及胶原酶消化法所得脂肪干细胞的成骨分化能力。方法分别采用组织块贴壁法及胶原酶消化法原代培养小鼠脂肪干细胞,观察细胞形态及细胞增殖情况,并进行细胞鉴定,比较两者成骨分化能力。结果两种方法所得脂肪干细胞在形态学上无差异,培养至第3代,细胞生长状态良好,生物学特性稳定,CD44、Sca-1表达率>90%,CD31表达率<10%,具有较好的成脂、成骨分化能力;组织块贴壁法培养所得脂肪干细胞增殖率高于胶原酶消化法(P<0.05);组织块贴壁法培养所得脂肪干细胞矿化结节、ALP活性及钙离子浓度高于胶原酶消化法(P <0.05)。结论组织块贴壁法相对胶原酶消化法在脂肪干细胞培养及成骨诱导分化过程中更具优势。

Objectives To investigate the influence of different culture methods on osteogenic differentiation of adipose-derived stem cells.Methods Adipose-derived stem cells(ASCs)were cultured by tissue cultivation(group A)or collagenase digestion method(group B).Morphology and cell proliferation of ASCs were observed.ASCs were identified,and osteogenic differentiation capacity was compared.Results No morphological differences were found among groups.ASCs grew well with stable biological property.Expression rate of CD44 and Sca-1 is greater than 90%,and the expression rate of CD31 is less than 10%in the third generation.Both cells were presented with fine adipogenic and osteogenic differentiation capacity.The proliferation rate of ACSs in group A was significantly increased than that in group B(P<0.05).Furthermore,mineralized nodule,ALP activity and concentration of calcium ion of ASCs in group A were enhanced than those in group B(P<0.05).Conclusions Tissue cultivation method exerts better advantages in ACSs culture and osteogenic differentiation compared with collagenase digestion method.