无血清培养脐带间充质干细胞的研究

发布时间：2018-08-25 16:36

【摘要】：本研究观察无血清培养液培养的人脐带间充质干细胞(UC-MSC),并比较与含10%胎牛血清培养液培养的人脐带间充质干细胞的异同。采集正常足月剖宫产胎儿脐带,用MesenCult-XF无血清培养液或含10%胎牛血清培养液培养。观察不同培养液培养的间充质干细胞细胞的形态、免疫表型、细胞周期、增殖分化潜能及对混合淋巴细胞反应的抑制作用。结果表明,采用无血清MesenCult湟-XF培养液培养的MSC平均传代倍增6.57±0.7倍,含血清培养液培养的MSC平均传代倍增4,59±0.45倍(P0.05);两种培养液培养的MSC均表达CD44、CD90、CD73、CD105抗原,不表达CD31、CD45、HLA-DR及CD34等抗原,表达程度无明显统计学差异;无血清培养液培养的MSC(65±5.2)%均为G o/G1期细胞,含血清培养液培养的MSC(62±3.1)%为G o/G1期细胞(P0.05);两种培养液培养的人脐带MSC均可向成脂细胞、成骨细胞分化;无血清培养液培养的脐带MSC按1 000、5 000、10 000、20 000个细胞/孔接种密度与反应细胞和刺激细胞共培养的每分钟闪烁值(CPM)分别为(6.43±0.47)×104、(4.30±0.38)×104、(1.97±0.13)×104和(0.24±0.03)×104,含血清培养液培养的MSC与不同密度的混合淋巴细胞共培养的CPM值分别为(7.85±0.07)×104、(5.64±0.12)×104、(3.09±0.18)×104和(1.73±0.05)×104。结论:无血清培养液培养的细胞均为MSC,有传代增殖潜能,传代可达临床治疗MSC细胞量,并可避免异种蛋白致敏。
[Abstract]:Human umbilical cord mesenchymal stem cells (UC-MSC) cultured in serum-free medium were observed and compared with human umbilical cord mesenchymal stem cells cultured in 10% fetal bovine serum culture medium. Umbilical cord of normal full-term caesarean section was collected and cultured with MesenCult-XF serum-free culture medium or 10% fetal bovine serum culture medium. The morphology, immunophenotype, cell cycle, proliferation and differentiation potential of mesenchymal stem cells cultured in different culture medium were observed. The results showed that the average passage doubling of MSC in serum-free MesenCult Huang XF medium was 6.57 卤0.7-fold, and that of MSC cultured in serum-containing medium was 40.59 卤0.45fold (P0.05). The MSC in both cultures expressed CD44,CD90,CD73,CD105 antigens, but did not express CD31,CD45,HLA-DR and CD34 antigens. The expression of MSC in serum-free medium (65 卤5.2)% was G o/G1 phase cells and MSC (62 卤3.1)% in serum-containing medium was G o/G1 phase cells (P0.05). Human umbilical cord MSC could differentiate into adipogenic cells and osteoblasts. The (CPM) of umbilical cord MSC cultured in serum-free medium was (6.43 卤0.47) 脳 10 ~ 4, (4.30 卤0.38) 脳 10 ~ 4, (1.97 卤0.13) 脳 10 ~ 4 and (0.24 卤0.03) 脳 10 ~ (4), respectively. The (CPM) of the co-culture of the cells / well and response cells and stimulating cells was (6.43 卤0.47) 脳 10 ~ (4), (1.97 卤0.13) 脳 10 ~ 4 and (0.24 卤0.03) 脳 10 ~ 4 respectively. The CPM values of co-cultured mixed lymphocytes were (7.85 卤0.07) 脳 10 ~ 4, (5.64 卤0.12) 脳 10 ~ 4, (3.09 卤0.18) 脳 10 ~ 4 and (1.73 卤0.05) 脳 10 ~ 4, respectively. Conclusion: all the cells cultured in serum-free medium have the potential to proliferate through passage, which can reach the clinical treatment of MSC cells and avoid the allergy of xenogeneic proteins.
【作者单位】：苏州大学附属儿童医院血液肿瘤科;苏州大学附属第一医院江苏省血液研究所卫生部血栓与止血重点实验室;
【基金】：江苏省卫生厅面上项目(H201212) 苏州市科技发展计划项目(SS0718) 江苏省临床医学中心血液病学开放课题编号(KF200943)
【分类号】：R329.28

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