Expressions of PDGF receptor alpha, c‐Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells
Expressions of PDGF receptor alpha, c‐Kit and Flk1 genes clustering in mouse chromosome 5 define distinct subsets of nascent mesodermal cells
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DOI：
10.1046/j.1440-169x.1997.t01-5-00009.x
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发表时间：
1997-12
期刊：
影响因子：
4.6
通讯作者：
H. Kataoka;N. Takakura;S. Nishikawa;K. Tsuchida;H. Kodama;T. Kunisada;W. Risau;T. Kita;S. Nishikawa
中科院分区：
文献类型：
--
作者：
H. Kataoka;N. Takakura;S. Nishikawa;K. Tsuchida;H. Kodama;T. Kunisada;W. Risau;T. Kita;S. Nishikawa
在原肠胚中，在分化成特定谱系之前会产生各种类型的细胞。原肠胚形成的小鼠胚胎的中胚层代表了一组这样的中间细胞。 PDGF 受体 α (PDGFRα)、c-Kit 和胎儿肝激酶 1 (Flk1) 在原肠胚形成后胚胎的独特中胚层衍生物中表达。通过使用针对这三种受体的单克隆抗体进行整体免疫染色来检查它们在原肠胚形成过程中的表达。这些抗体对原肠胚中不同的中胚层子集进行了染色。头折阶段胚胎的流式细胞术显示，Flk1+ 中胚层细胞可以根据 c-Kit 表达水平进一步分类。为了检查造血细胞分化是否从 Flk1+ 中胚层开始的可能性，将胚胎干 (ES) 细胞培养在 OP9 或 PA6 基质细胞层上；前者而非后者支持ES细胞体外造血。从分化第 3 天起，在造血细胞出现之前，仅在 OP9 细胞层上检测到 Flk1+ 细胞。因此，Flk1+细胞是体外ES细胞分化为造血细胞所必需的。结果表明，这三种受体酪氨酸激酶可用于定义和分选来自胚胎或体外培养的 ES 细胞的中胚层细胞亚群。
In gastrulating embryos, various types of cells are generated before differentiation into specific lineages. The mesoderm of the gastrulating mouse embryo represents a group of such intermediate cells. PDGF receptor alpha (PDGFRα), c‐Kit and fetal liver kinase 1 (Flk1) are expressed in distinctive mesodermal derivatives of post‐gastrulation embryos. Their expressions during gastrulation were examined by whole mount immunostaining with monoclonal antibodies against these three receptors. The antibodies stained different mesodermal subsets in gastrulating embryos. Flow cytometry of head fold stage embryos revealed that Flk1+ mesodermal cells could be further classified by the level of c‐Kit expression. To examine the possibility that hematopoietic cell differentiation is initiated from the Flk1+ mesoderm, embryonic stem (ES) cells were cultured on the OP9 or PA6 stromal cell layer; the former but not the latter supported in vitro hematopoiesis from ES cells. Flk1+ cells were detected only on the OP9 cell layer from day 3 of differentiation before the appearance of hematopoietic cells. Thus, Flk1+ cells will be required for in vitro ES cell differentiation into hematopoietic cells. The results suggest that these three receptor tyrosine kinases will be useful for defining and sorting subsets of mesodermal cells from embryos or in vitro cultured ES cells.