脑瘫患者骨髓间充质干细胞传代过程中细胞表型和所分泌细胞因子变化特点

摘要

目的 分析脑瘫患者骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSC)的基本生物学特性,观察其在传代培养过程中的表型和所分泌的因子变化,为临床应用提供理论依据.方法 密度梯度离心法提取脑瘫患者的BMSCs,采用人淋巴细胞分离液分离BMSCs,在相同条件下观察各代细胞形态、生长情况,检测各代细胞的表型和分泌的细胞因子.结果 BMSCs在第3至第10代,CD29、CD44、CD106等表型的表达变化无统计学差异,CD54的表达变化最大,在第9代时最高.CD14、CD34、CD45和HLA-DR等细胞表型在各代骨髓间充质干细胞中表达均<5%;NGF、NT3、VCAM、ICAM-1、VEGF、BDNF等细胞因子的浓度变化无统计学差异,PGE2的变化需进一步研究予以确认.结论 在体外培养的条件下,8代以前的细胞生长形状稳定,可作为组织工程等临床应用的良好对象.%Objective To investigate the characteristic change of phenotype and cytokine of bone marrow derived mesenchymal stem cells (BMSCs) in cerebral palsy patients. Methods BMSCs were isolated by density gradient centrifugation from lydroxypropylmethyl cellulose. The morphology,growth,phenotype and cytokine of each passage cell were investigated. Results The phenotype of CD29,CD44, CD106 was not significantly different in each passage cell. The phenotype of CD54 was highest at P9. The expression of the phenotype of CD14,CD34,CD45 and HLA-DR was below 5％ in each passage cell. The changes of concentration of NGF, NT3, VCAM,ICAM - 1, VEGF, BDNF were not significant across the passage cells. The changes of PGE2 need to be further studied. Conclusions The state of BMSCs before the 8th passage is stable,and can be a valuable subject for basic research and clinical application.