低温冻存对人脐带间充质干细胞生物学特性的影响

Effect of cryopreservation on biological features of human umbilical cord mesenchymal stem cells

目的比较人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,UC-MSCs)冻存前与复苏后的生物学特性,为规模化储备UC-MSCs提供试验支持。方法采用胶原酶消化法从脐带中分离UC-MSCs,贴壁培养传代,将第3代UC-MSCs利用程控降温仪冷冻,置于-196℃液氮中冻存6个月后复苏,比较冻存前和复苏后UC-MSCs的细胞形态、生长曲线、免疫表型及多向分化潜能等生物学特性。结果复苏后UC-MSCs仍呈成纤维样形态生长,生长曲线与冻存前相似;免疫表型仍高表达CD73、CD90、CD105,不表达CD34、CD45、CD40、CD80、CD86、CD154、HLA-DR;在特定的体外诱导条件下,仍可以向骨、脂肪、软骨分化。结论冻存复苏后UC-MSCs的生物学特性仍保持稳定。

Objective To provide the experimental support for large-scale preservation of human umbilical cord mesenchymal stem cells（UC-MSC） by comparing their biological features before cryopreservation and after thawing.Methods Human UC-MSC were isolated by collagenase digestion and subcultured in vitro.The third passage human UC-MSC were frozen with a controlled rate freezer and thawed 6 months after they were stored in-196 ℃ liquid nitrogen.Their biological features were compared,including morphology,growth curve,immunophenotype and multi-directional differentiation potential before cryopreservation and after thawing.Results The human UC-MSC grew in a fibroblastic morphology after thawing with a similar growth curve to that before cryopreservation.The immunophenotype of human UC-MSC highly expressed the CD73,CD90,CD105 but not the CD34,CD45,CD40,CD80,CD86,CD154,HLA-DR.Human UC-MSC could differentiate into osteocytes,adipocytes and chondrocytes under special in vitro induction conditions.Conclusion Human UC-MSC can maintain their biological features after cryopreservation and thawing.