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【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。
【Objective】 To compare the biological characteristics of three types of mesenchymal stem cells (MSC) derived from human umbilical cord blood, neonatal umbilical cord blood and adult bone marrow in order to provide an experimental basis for selecting the cell source for clinical treatment. 【Method】 Three kinds of MSC were isolated and cultured by L-DMEM. Cell phenotype, cell cycle and apoptosis were analyzed by flow cytometry. Osteogenic and adipogenic inducing fluids were induced to differentiate into cells, respectively, carrying green fluorescent protein The recombinant adenovirus was transfected into MSCs. Fluorescent microscopy and flow cytometry were used to detect the expression of foreign genes in the three kinds of cells. 【Results】 MSCs could be isolated and cultured in all three samples, but the positive rate of neonatal cord blood was the lowest (29.17%, 7/24). The cell morphology, surface antigen expression and differentiation in vitro were similar among the three samples The primary culture time of cord blood MSCs was about 30 days, and the percentage of cells in G0 / G1 phase was higher than that in adult bone marrow. All of the 3 cells could be transfected to express foreign genes, but the highest expression rate of embryonic cord MSC was (56.32 ± 3.28)%. 【Conclusion】 MSCs can be isolated and cultured in embryonic, neonatal cord blood and adult bone marrow; mid-term embryonic cord blood MSCs are suitable for fetal intrauterine gene transfer / therapy (IUGT) target cells; adult bone marrow MSCs in tissue engineering and cell therapy , Gene therapy and other fields have a broad clinical application prospects; neonatal cord blood MSC clinical application needs further study.