Ward Christopher M, Barrow Katie M, Stern Peter L
Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK.
Exp Cell Res. 2004 Feb 15;293(2):229-38. doi: 10.1016/j.yexcr.2003.10.017.
Mouse embryonic stem (ES) cells are isolated from the inner cell mass (ICM)/epiblast of preimplantation embryos and are widely used in cell differentiation studies. We have previously observed differences in transcript and antigen expression following differentiation of ES cells lines in vitro. We have investigated this further by comparing the differentiation characteristics of five independently derived ES cell lines cultured and differentiated under defined conditions. Undifferentiated ES cell lines exhibited similar morphology and antigen/transcript marker expression. However, upon differentiation in monolayer culture by LIF withdrawal, only two of the lines expressed similar germ layer transcript profiles, and these were significantly altered compared to differentiation in serum-supplemented media. Neurofilament-68k was the only transcript marker common to all cell lines, however, induction of neuroectoderm lineages using 1 microM all-trans retinoic acid (RA) resulted in significant variations in cell number and morphology between the lines. Furthermore, neurons were only formed from clones of the two cell lines that exhibited similar transcript profiles, although the morphology was different between the two. We conclude that the independent ES cell lines in this study differ in their response to alterations in culture conditions in vitro, and the use of an appropriate cell line enables relatively homogeneous neuronal populations to be achieved in monolayer culture under defined conditions.
小鼠胚胎干细胞（ES细胞）是从植入前胚胎的内细胞团（ICM）/上胚层中分离出来的，广泛应用于细胞分化研究。我们之前观察到ES细胞系在体外分化后转录本和抗原表达存在差异。我们通过比较在特定条件下培养和分化的五个独立来源的ES细胞系的分化特性，对此进行了进一步研究。未分化的ES细胞系表现出相似的形态以及抗原/转录本标志物表达。然而，在通过撤除白血病抑制因子（LIF）进行单层培养分化时，只有两个细胞系表达相似的胚层转录本谱，并且与在添加血清的培养基中分化相比，这些转录本谱有显著改变。神经丝蛋白-68k是所有细胞系共有的唯一转录本标志物，然而，使用1微摩尔全反式维甲酸（RA）诱导神经外胚层谱系会导致各细胞系之间细胞数量和形态出现显著差异。此外，神经元仅从表现出相似转录本谱的两个细胞系的克隆中形成，尽管两者的形态有所不同。我们得出结论，本研究中的独立ES细胞系对体外培养条件变化的反应不同，并且使用合适的细胞系能够在特定条件下的单层培养中获得相对均一的神经元群体。
