论文部分内容阅读
目的:观察脐带血清等4种不同培养体系体外培养人骨髓间充质干细胞(BM-MSCs)的效果,建立一种适合临床移植需要的人BM-MSCs培养方法。方法:从20例新鲜成人骨髓穿刺液中分离BM-MSCs,分别用含10%脐带血清、胎牛血清(FCS)、成人血清的DMEM/F12培养基和MesenCult培养基培养,流式细胞仪检测细胞表面抗原,成骨诱导体系及脂肪诱导体系分别诱导各组BM-MSCs定向分化。结果:采用4种培养体系都能从骨髓液中分离培养出BM-MSCs,脐带血清和MesenCult组培养的BM-MSCs在增殖数量和速度上相似,而优于FCS和AB型血清组。脐带血清和MesenCult组培养的BM-MSCs表达CD29、CD73、CD105的阳性率比FCS和AB型血清组高(均P<0.05),而CD31阳性率低于FCS和AB型血清组(P<0.05),脐带血清和MesenCult组培养的BM-MSCs诱导为成骨细胞和脂肪细胞阳性率高于FCS和AB型血清组(P<0.05)。结论:脐带血清与MesenCult组培养的BM-MSCs的纯度和体外诱导分化能力相似,优于FCS和AB型血清组,脐带血清适合临床应用。
OBJECTIVE: To observe the effects of umbilical cord serum and other 4 culture systems on human bone marrow mesenchymal stem cells (BM-MSCs) in vitro and to establish a suitable culture method for human BM-MSCs for clinical transplantation. Methods: BM-MSCs were isolated from 20 fresh adult bone marrow aspirates and cultured in DMEM / F12 medium containing 10% cord blood serum, fetal bovine serum (FCS), adult serum and MesenCult medium, respectively. Flow cytometry Cell surface antigens, osteogenic and adipogenic systems induced the differentiation of BM-MSCs in each group. Results: BM-MSCs could be isolated and cultured from bone marrow fluid by using four culture systems. BM-MSCs cultured in cord blood serum and MesenCult group were similar in quantity and speed of proliferation but superior to FCS and AB type serum groups. The positive rates of CD29, CD73 and CD105 expression in cord blood serum and MesenCult group were higher than those in FCS and AB group (all P <0.05), while the positive rate of CD31 was lower than those in FCS and AB group (P <0.05 ). The positive rates of osteoblasts and adipocytes induced by BM-MSCs cultured in cord blood serum and MesenCult group were higher than those in FCS and AB type serum groups (P <0.05). Conclusion: The purity of BM-MSCs cultured in cord blood serum and MesenCult group is similar to that of FCS and AB-type serogroups in vitro, and the cord blood serum is suitable for clinical application.