期刊文献+

形成拟胚体细胞数量对小鼠胚胎干细胞心肌细胞及内皮细胞分化的影响

Effect of the Starting Number of Cells in Embryoid Bodies on Cardiac and Endothelial Differentiation of Murine Embryonic Stem Cells
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摘要 目的:探讨形成拟胚体(EBs)起始小鼠胚胎干细胞(ESCs)数量对心肌细胞及内皮细胞分化的影响。方法:将形成EBs起始ESCs数量为400,1 000,4 000的3个组(HD400,HD1000,HD4000)通过悬滴法促进其分化。免疫荧光检测TnT及CD31的表达;在不同时间点计算三个组搏动EBs百分比;RT-PCR检测Nkx2.5,GA-TA4,β-MHC,flk-1,CD31,tie-2基因表达水平。结果:通过悬滴法形成的EBs能够出现自发性搏动并且有TnT及CD31表达;HD1000组和HD4000组相比HD400组有更高的搏动EBs百分比及Nkx2.5,GATA4,β-MHC基因表达水平;HD400组相比HD1000组和HD4000组有更高的flk-1,CD31,tie-2基因表达水平(P<0.05)。结论:形成EBs起始ESCs数量是影响心肌细胞和内皮细胞分化的一个重要因素,起始细胞数量多则心肌细胞分化程度高,数量少则内皮细胞分化程度高。 Objective: To investigate the effect of the starting number of murine embryonic stem cells (ESCs) in embryoid bodies (EBs) on the cardiac and endothelial differentiation. Methods: Hang- ing drop culture was used to induce ESCs differentiating into cardiomyocytes and endothelial cells. The groups of the EBs formed from 400, 1 000, and 4 000 ESCs were referred to as HD400, HD1000 and HD4000, respectively. TnT and CD31 were detected by immunocytochem- istry. The percentage of beating EBs was calculated at different time points. The mRNA expres- sion of cardiac specific gene Nkx2.5, GATA4, β-MHC and endothelial specific gene flk-1, CD31, and tie-2 were detected by RT-PCR. Results. Spontaneously beating EBs were positively stained with TnT or CD31. The percentage of beating EBs and Nkx2.5, GATA4,β-MHC gene expres- sion were higher in the HD4000 group and HD1000 group compared with HD400 group. Moreo- ver, the HD400 group had higher gene expression of flk-1, CD31, tie-2. Conclusion. Our resultsindicate that the starting number of ESCs in EBs is important in regulation of the cardiac and endothe- lial differentiation of ESCs. The cardiac differentiation is increased in the EBs by a larger number of ESCs, while the endothelial differentiation is increased in the EBs by a smaller number of ESCs.
出处 《武汉大学学报(医学版)》 CAS 北大核心 2013年第2期157-160,165,共5页 Medical Journal of Wuhan University
基金 国家自然科学基金资助项目(编号:81200119) 中央高校基本科研业务费专项资金(编号:121007)
关键词 胚胎干细胞 心肌细胞 内皮细胞 分化 悬滴法 Embryonic Stem Cell Cardiomyocyte Endothelial Cell Differentiation Hang- ing Drop Method
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参考文献9

  • 1Chen M, I.in YQ, Xie SI., et al. Mitogen-activated protein kinase in endothelin-I induced cardiac differentiation of mouse embryonic stem cells[J]. J Cell Biochem, 2010, 111(6)-1 619-1 628.
  • 2Abranches E,Bekman E, Henrique D, et al. Expansion and neural differentiation of embryonic stem cells in adherent and suspension cultures [J]. Biotechnol Lett, 2003, 25 (9):725-730.
  • 3Chen M, l.in YQ, Xie SI., et al. Enrichment of Cardiac Differentiation of Mouse I-2mbryonic Stem Cells hy optimizing the Hanging Drop Method [J]. Biotechnol I.ett, 2011, 33(4)..853 858.
  • 4Boheler KR, Czyz J, Tweedie D, et al. Differentiation of pluripotent embryonic stem cells into cardiomyocytes [J]. Circ Res, 2002, 91 (3) :189-201.
  • 5Chen K, Wu L, Wang ZZ. Extrinsic regulation of cardiomyocyte differentiation of embryonic stem cells [J-. J Cell Biochem, 2008, 104(1):119 128.
  • 6Park J, Cho CH, Parashurama N, el al. Microfabrication-based modulation of embryonic stem cell differentiation[J]. Lab Chip, 2007, 7 (8):1 018- 1 028.
  • 7Bratt-Leal AM, Carpenedo RL, McDevitt TC. Engineering the embryoid body microenvironment to direct embryonic stem cell differentiadon [J]. Biotechnol Prog, 2009, 25: 43-51.
  • 8Hwang YS, Chung BG, Ortmann D, et al. Microwell- mediated control of embryoid body size regulates embryonic stem cell fate via differential expression of WNT5a and WNT11 [J]. Proc Natl Aead Sci USA, 2009, 106:16 978-16 983.
  • 9Naito AT, Shiojima I, Akazawa H, et al. Developmental stage-specific biphasic roles of Wnt/beta-eatenin signaling in cardiomyogenesis and hematopoiesis[J]. Proc Natl Acad Sei USA, 2006, 103:19 812-19 817.

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