Lentiviral rescue of vascular endothelial growth factor receptor-2 expression in Flk1-/- embryonic stem cells shows early priming of endothelial precursors
Lentiviral rescue of vascular endothelial growth factor receptor-2 expression in Flk1-/- embryonic stem cells shows early priming of endothelial precursors
复制标题
DOI：
10.1634/stemcells.2007-0397
复制
发表时间：
2007-01-01
期刊：
影响因子：
5.2
通讯作者：
Claesson-Welsh, Lena
中科院分区：
文献类型：
--
作者：
Li, Xiujuan;Edholm, Dan;Claesson-Welsh, Lena
血管内皮生长因子 (VEGF) 家族及其受体对于血管发育和血管维护以及血管生成（新血管的形成）非常重要。 VEGF 受体 2（VEGFR-2；在小鼠中称为 Flk-1）的缺失会导致体内血管和造血发育停滞。我们使用慢病毒转导来重建 flk1-/- 胚胎干 (ES) 细胞中的 VEGFR-2 表达。 VEGF 诱导的血管生成和萌芽血管生成在体外分化为 EB 的转导 ES 培养物中得到挽救。尽管转基因在多能干细胞中表达并且在分化过程中缺乏谱系限制，但内皮募集的程度与野生型 EB 中的相似。 flk1-/- ES 细胞中 VEGFR-2 的重建仅允许预先定型的前体细胞分化为能够组织成血管结构的功能性内皮细胞。创建了由野生型 ES 细胞与 flk1-/- ES 细胞或重建的表达 VEGFR-2 的 ES 细胞混合组成的嵌合 EB 培养物。在嵌合培养物中，flk1-/-内皮前体被排除在野生型血管结构之外，而重组的表达VEGFR-2的前体与野生型内皮细胞整合在一起形成嵌合血管。我们得出结论，内皮前体的成熟以及组织成血管结构需要 VEGFR-2 的表达。
The vascular endothelial growth factor ( VEGF) family and its receptors are important for vascular development and maintenance of blood vessels, as well as for angiogenesis, the formation of new vessels. Loss of VEGF receptor-2 (VEGFR-2; designated Flk-1 in mouse) results in arrest of vascular and hematopoietic development in vivo. We used lentiviral transduction to reconstitute VEGFR-2 expression in flk1-/- embryonic stem (ES) cells. VEGF-induced vasculogenesis and sprouting angiogenesis were rescued in transduced ES cultures differentiating in vitro as EBs. Although the transgene was expressed in the pluripotent stem cells and lacked linage restriction during differentiation, the extent of endothelial recruitment was similar to that in wild-type EBs. Reconstitution of VEGFR-2 in flk1-/- ES cells allowed only precommitted precursors to differentiate into functional endothelial cells able to organize into vascular structures. Chimeric EB cultures composed of wild-type ES cells mixed with flk1-/- ES cells or reconstituted VEGFR-2expressing ES cells were created. In the chimeric cultures, flk1-/- endothelial precursors were excluded from wild-type vessel structures, whereas reconstituted VEGFR-2-expressing precursors became integrated together with wild-type endothelial cells to form chimeric vessels. We conclude that maturation of endothelial precursors, as well as organization into vascular structures, requires expression of VEGFR-2.