Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies
Characterization of embryoid bodies of mouse embryonic stem cells formed under various culture conditions and estimation of differentiation status of such bodies
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DOI：
10.1263/jbb.104.294
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发表时间：
2007-10-01
影响因子：
2.8
通讯作者：
Kurosawa, Hiroshi
中科院分区：
文献类型：
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作者：
Koike, Mikiko;Sakaki, Shujiro;Kurosawa, Hiroshi
对小鼠胚胎干细胞（ES细胞）在不同培养条件下形成的不同类型的拟胚体（EB）进行了基因表达谱的表征，以估计胚体的分化状态。典型标记的基因表达（即，在各种类型的EB中定量分析了加塔-4、加塔-6、甲状腺素运载蛋白[TTR]、甲胎蛋白[AFP]、Nkx 2.5和α-肌球蛋白重链[α-MHC]），并且对于每种EB，图示了这些标记基因的基因表达模式。基因表达谱能准确反映EB的分化状态。基因表达模式表明，Nkx2.5和α-MHC基因在低粘附96孔板中由1000个ES细胞形成的EB中高度表达。通过将EB转移到附着培养物中，在EB的生长中更有效地产生心肌细胞。当我们将种子细胞数量从1000增加到4000个ES细胞时，基因表达模式发生了变化，即TTR和AFP基因的表达水平增加，而Nkx2.5和α-MHC基因的表达水平降低，并且分化趋势从心肌样向内脏卵黄囊样结构形成转变。
Various types of embryoid body (EB) that were formed from mouse embryonic stem (ES) cells under various culture conditions were characterized in terms of gene expression pattern to estimate the differentiation status of the bodies. The gene expression of typical markers (i.e., GATA-4, GATA-6, transthyretin [TTR], alpha-fetoprotein [AFP], Nkx2.5, and alpha-myosin heavy chain [alpha-MHC]) was quantitatively analyzed in various types of EB, and the gene expression pattern of those marker genes was graphically shown for each EB. The gene expression pattern accurately represented the differentiation status of the EBs. The gene expression pattern indicated that the Nkx2.5 and alpha-MHC genes were highly expressed in the EBs formed from 1000 ES cells in a low-adherence 96-well plate. By transferring the EBs into an attachment culture, cardiomyocytes were more efficiently generated in the outgrowth of the EBs. When we increased the seeding cell number from 1000 to 4000 ES cells, the gene expression pattern changed, that is, the expression levels of the TTR and AFP genes increased, whereas those of the Nkx2.5 and alpha-MHC genes decreased, and the trend of differentiation changed from cardiomyogenesis to visceral yolk-sac-like structure formation.