人中期胚胎、新生儿脐血及成人骨髓间质干细胞基本生物学特性的比较

Comparative Study of Basic Biological Characteristics of Mesenchymal Stem Cells Derived from Human Fetal, Newborn Cord Blood, and Adult Bone Marrow

【目的】比较人中期胚胎脐血、新生儿脐血以及成人骨髓3种来源的间质干细胞(MSC)生物学特性,为临床治疗选择细胞来源提供实验依据。【方法】采用L-DMEM培养液分离培养3种来源MSC,流式细胞仪进行细胞表型、细胞周期、细胞凋亡分析,成骨、成脂诱导液分别诱导细胞分化,携带绿色荧光蛋白报告基因的重组腺病毒转染MSC,荧光显微镜和流式细胞仪检测3种细胞表达外源基因特性。【结果】3种来源标本中均可分离培养出MSC,但新生儿脐血中阳性获得率最低,为29.17%(7/24);3者细胞形态、表面抗原表达、体外分化特性相似,但2种脐血MSC原代培养时间长达30d左右,处于G0/G1期细胞百分比大于成人骨髓;3者均能转染表达外源基因,但胚胎脐血MSC中表达率最高,达(56.32±3.28)%。【结论】胚胎、新生儿脐血以及成人骨髓中均可分离培养出MSC;中期胚胎脐血MSC适用于胎儿自体宫内基因转移/治疗(IUGT)靶细胞;成人骨髓MSC在组织工程、细胞治疗、基因治疗等领域具有广阔的临床应用前景;而新生儿脐血MSC的临床应用有待进一步研究。

ObjectiveTo investigate the differences of biological characteristics of mesenchymal stem cells(MSC) derived from human second-trimester fetal, newborn cord blood, and adult bone marrow.MethodsL-DMEM medium was used to cultivate MSC. The immunophenotype, cell cycle, and cell apoptosis were analyzed by flow cytometry. Adipogenic and osteogenic mediums were used to assess the differentiation ability of the cells. Adenovirus vector delivered green fluorescent protein gene (Ad-GFP) was used to transfect the cells. ResultsFetal, newborn cord blood, and bone marrow derived cells cultured in L-DMEM gave rise to adherent cells with fibroblast-like morphology, but the positive rate in newborn cord blood was the lowest as 29.17%(7/24). These cells were similar in cell morphology, antigenic phenotype, differentiation potential, and cell apoptosis rates. The primary culture time of cord blood derived MSC was longer than that of bone marrow derived cells. The percentage of G0/G1 cells in cord blood was higher than that in bone marrow. These cells transfected and expressed the transgene, but the fetal derived cells expressed at the highest level (56.32±3.28%).ConclusionHuman second-trimester fetal cord blood MSC is an promising target cell in fetal uterus gene transfer/therapy; the adult bone marrow derived MSC is useful in bioengineering, cell and gene therapy; whereas the application of newborn cord blood MSC need more researching.