人脐带间充质干细胞冻存复苏后生物学性状研究

Biological Characteristics of Cryopreserved Human Umbilical Cord Mesenchymal Stem Cells

目的观察冻存前与冻存复苏后人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCM-SCs)生物学特性的变化。方法从我院产科健康剖宫产产妇捐献的脐带中分离得到hUCMSCs,观察其生长情况,检测其表面抗原,并将第3代hUCMSCs采用梯度冷冻技术冻存6个月,比较冻存前及冻存复苏后hUCMSCs的形态、增殖、染色体、免疫表型情况及多向分化能力。结果冻存前及冻存复苏后,hUCMSCs形态无明显差异,均呈典型梭形,贴壁生长;荧光强度、平均存活率比较差异无统计学意义(P>0.05);生长曲线、免疫表型相似。冻存复苏后的hUCMSCs染色体检查未见异常,有向脂肪细胞分化的能力。结论 hUCMSCs经冻存复苏后生物学特性仍保持稳定。

Objective To investigate biological characteristics of the human umbilical cord mesenchymal stem cells(hUCMSCs).Methods hUCMSCs were isolated from donated umbilical cord in women after cesarean section and the cell growth was observed and its surface antigen was detected.The three-passage hUCMSCs were preserved in liquid nitrogen by lowering the temperature gradually.Six months later,cells were thawed,the morphology,proliferation,chromosome,immunophenotype and differentiation of hUCMSCs were investigated and compared with those of hUCMSCs before cryopreservation.Results There was no significant difference of morphology between pre-cryopreserved hUMSCs and the ones after resuscitation.It was obvious that all hUMSCs were spindle-shaped and showed adherence growth characteristic before and after cryopreservation.The cell growth curves,inmunophenotype of hUMSCs were also similar before and after cryopreservation.Fluorescence intensity and average survival rate showed no significant differences(P>0.05).Chromosome karyotype didn′t show any significant abnormality,and hUCMSCs had the ability of differentiating into adipocytes.Conclusion hUCMSCs derived from human umbilical cord still maintain their biological characteristics after cryopreservation and resuscitation.