摘要
目的探讨冻存和复苏对人脐带间充质干细胞(mesenchymal stem cells,MSCs)的形态、表型和向神经细胞分化的潜能的影响。方法在无菌条件下采集脐带组织,以酶消化和贴壁培养法获得人脐带MSCs。对不同代人脐带MSCs进行冻存和复苏,以相差显微镜观察期形态学变化,以流式细胞仪检测表型变化,以神经分化方案进行诱导并以免疫荧光法检测各种神经标志物的表达。结果不同代冻存后复苏的人脐带MSCs仍呈均匀一致的长梭形贴壁生长,表达MSCs标志物CD13、CD29、CD44和CD90,但不表达造血细胞标志物CD14、CD31、CD34和CD45。经神经分化诱导后,仍可分化为NSE阳性的神经元样细胞、GFAP阳性的星形胶质细胞样细胞和MBP阳性的少突胶质细胞样细胞。结论冻存和复苏后人脐带MSCs的形态、表型和神经分化潜能均保持稳定,可作为快速获得人脐带MSCs的方法。
Objective To investigate the effects of cryopreservation on the neural differentation potential of human umbilical cord mesenchymal stem cells （bUC-MSCs）. Methods The umbilical cords were collected under sterile condi- tions, and MSCs were isolated using enzyme digestion methods and adherent culture. After frezing-thawing, the morphology of umbilical cord MSCs were observed under phase-contrast microscope, the phenotypes were analysed by flow cytometry, and the neural differentiation potentials were confirmed after neural induction and immunofluorescence staining. Results The umbilical cord MSCs remained their adherent characteristic, spindle-like morphology, mesencbymal phenotypcs （expres- sion of CD13, CD29, CIM4 and CI）90,but without expression of CD14, CD31 ,CD34 and CD45 ）and neural differention po- tential into NSE stained neuron-like cells, GFAP stained astrocyte-like cells and MBP stained oligodendrocyte-like cells. Conclusions The morphology, pbenotypes and neural differentiation potential remained relatively stable after cryoprescrva- tion, paving way for direct and convenient application of pre-frozened bUC-MSCs.
出处
《中风与神经疾病杂志》
CAS
CSCD
北大核心
2013年第11期964-967,共4页
Journal of Apoplexy and Nervous Diseases
基金
国家自然科学基金项目(81001009)
关键词
间充质干细胞
脐带
冻存
神经分化
Mesenchymal stem cell
Umbilical cord
Cryopreservation
Neural differentaition
