VEGF and IHH rescue definitive hematopoiesis in Gata-4 and Gata-6-deficient murine embryoid bodies.
VEGF and IHH rescue definitive hematopoiesis in Gata-4 and Gata-6-deficient murine embryoid bodies.
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DOI：
10.1016/j.exphem.2009.05.011
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发表时间：
2009-09
影响因子：
2.6
通讯作者：
Yoder, Mervin C.
中科院分区：
文献类型：
--
作者：
Pierre, Monique;Yoshimoto, Momoko;Huang, Lan;Richardson, Matthew;Yoder, Mervin C.
小鼠胚胎干细胞可分化为拟胚体，拟胚体可作为模拟胚胎卵黄囊造血过程的体外模型。缺乏加塔-4或加塔-6的ES细胞的分化导致具有破坏的内脏内胚层（VE）的EB。虽然缺乏维生素E对体内造血有不利影响，但目前尚不清楚缺乏维生素E是否会影响EB的造血。因此，我们比较了加塔-4 null（G4 N）和加塔-6 null（G6 N）EB与野生型（WT）EB，以评估它们定型为造血细胞的能力。用细胞分选技术检测EB-VE形成，并分析内脏内胚层基因表达。使用集落形成测定评估在各种条件下培养的EB的造血祖细胞潜能。与WT EB相比，G4 N和G6 N EB中的嗜酸性红细胞、粒细胞-巨噬细胞和混合集落显著减少。血管内皮生长因子（VEGF）的表达和分泌也减少在G4 N和G6 N EB，作为VEGF生产的网站与VE一致。向EB培养物中添加外源性VEGF 165完全拯救了G4 N和G6 N EB中的永久集落形成细胞。这种拯救反应可以通过向EB培养物中加入可溶性Flk-1 Fc来阻断。类似地，向EB培养物中添加外源性印度刺猬（IHH）也以可逆的方式恢复了确定性造血的减少。这些结果表明，在G4 N和G6 N EB中VE的缺乏并不能阻止EB中定形祖细胞的出现。然而，缺乏VE的G4 N和G6 N EB中VEGF和IHH产生水平的降低减弱了永久性造血祖细胞扩增。
Murine embryonic stem (ES) cells can be differentiated into embryoid bodies (EBs), which serve as an in vitro model recapitulating many aspects of embryonic yolk sac hematopoiesis. Differentiation of ES cells deficient in either Gata-4 or Gata-6 results in EBs with disrupted visceral endoderm (VE). While lack of VE has detrimental effects on hematopoiesis in vivo, it is unclear whether lack of VE affects hematopoiesis in EBs. Therefore, we compared Gata-4 null (G4N) and Gata-6 null (G6N) EBs with wild-type (WT) EBs to assess their ability to commit to hematopoietic cells. EB VE formation was examined using cell sorting techniques and analysis visceral endoderm gene expression. Hematopoietic progenitor potential of EBs cultured under various conditions was assessed using colony forming assays. Definitive erythroid, granulocyte-macrophage, and mixed colonies were significantly reduced in G4N and G6N EBs compared to WT EBs. Vascular endothelial growth factor (VEGF) expression and secretion were also reduced in both G4N and G6N EBs, consistent with VE serving as a site of VEGF production. Addition of exogenous VEGF165, to EB cultures completely rescued definitive colony forming cells in G4N and G6N EBs. This rescue response could be blocked by addition of soluble Flk-1 Fc to EB cultures. Similarly, addition of exogenous Indian hedgehog (IHH) to EB cultures also recovers the diminishment in definitive hematopoiesis in a reversible manner. These results suggest that the absence of VE in G4N and G6N EBs does not prevent the emergence of definitive progenitors from EBs. However, the decreased level of VEGF and IHH production in VE devoid G4N and G6N EBs attenuates definitive hematopoietic progenitor cell expansion.
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影响因子：
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KELLER, G;KENNEDY, M;WILES, MV
通讯作者：
WILES, MV
影响因子：
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