江苏大学学报（医学版）2014，Vol.24Issue(3)：190-194,5.DOI:10.13312/j.issn.1671-7783.y140070
低温冻存对人脂肪间充质干细胞生物学特性的影响
Effect of cryopreservation on biological characteristics of human adipose-derived mesenchymal stem cells
摘要
Abstract
Objective:To provide the experimental support for establishing a low-temperature stem cell bank for future use,freeze-thawed human adipose-derived mesenchymal stem cells (hADSCs) were compared with fresh hADSCs in vitro.Methods:The hADSCs were isolated from abdomen epiploica adipose tissue by collagenase digestion and subcultured in vitro.The third passage of hADSCs was cryopreserved in liquid nitrogen for 1,3,and 6 months.Cell viability,proliferation potential,expression of cellular markers and multipotential differentiation were studied after thawing using trypan blue staining,MTT method,flow cytometry analysis,Oil Red O staining,von Kossa,and RT-PCR.Results:After thawing,cell survival rate was found to descend as the time of cryopreservation prolonged.MTT revealed high capability for the proliferation.Phenotypic studies revealed that hADSCs were positive for CD29,CD44,CD73,CD90,CD105,CD166 and negative for CD45,and HLA-DR.Functional analysis demonstrated these cells could be differentiated into adipocytes and osteoblasts in lineage-specific medium.RT-PCR results proved that adipogenic and osteogenic related genes could be expressed.No significant difference (P ＞ 0.05) was noted before and after the cryopreservation in cell viability,proliferation potential,expression of cellular markers and multipotential differentiation.Conclusions:The hADSCs could maintain acceptable viability,biological characteristics,and multipotential differentiation after cryopreservation.关键词
人脂肪间充质干细胞/冻存/生物学特性/干细胞库Key words
human adipose-derived mesenchymal stem cells/ cryopreservation/ biological characteristics/ stem cell bank分类
医药卫生引用本文复制引用
李兴天,李小战,马红,许辉,李淑,李遇梅..低温冻存对人脂肪间充质干细胞生物学特性的影响[J].江苏大学学报（医学版）,2014,24(3):190-194,5.基金项目
国家自然科学基金资助项目(30972663) （30972663）
