Abstract
RationaleInduced pluripotent stem (iPS) cells have emerged as a source of potentially unlimited supply of autologous endothelial cells (ECs) for vascularization. However, the regenerative function of these cells relative to adult ECs and ECs derived from embryonic stem (ES) cells is unknown. The objective was to define the differentiation characteristics and vascularization potential of Fetal liver kinase (Flk)1+ and Vascular Endothelial (VE)-cadherin+ ECs derived identically from mouse (m)ES and miPS cells. Methods and ResultsNaive mES and miPS cells cultured in type IV collagen (IV Col) in defined media for 5 days induced the formation of adherent cell populations, which demonstrated similar expression of Flk1 and VE-cadherin and the emergence of EC progenies. FACS purification resulted in 100% Flk1+ VE-cadherin+ cells from both mES and miPS cells. Emergence of Flk1+VE-cadherin+ cells entailed expression of the vascular developmental transcription factor Er71, which bound identically to Flk1, VE-cadherin, and CD31 promoters in both populations. Immunostaining with anti-VE-cadherin and anti-CD31 antibodies and microscopy demonstrated the endothelial nature of these cells. Each cell population (unlike mature ECs) organized into well-developed vascular structures in vitro and incorporated into CD31+ neovessels in matrigel plugs implanted in nude mice in vivo.ConclusionThus, iPS cell-derived Flk1+VE-cadherin+ cells expressing the Er71 are as angiogenic as mES cell-derived cells and incorporate into CD31+ neovessels. Their vessel forming capacity highlights the potential of autologous iPS cells-derived EC progeny for therapeutic angiogenesis.
Highlights
The transduction of fibroblast cells with transcription factors Nanog, Sox2, Oct4, Klf4, and c-Myc converts these cells into induced pluripotent stem cells [1,2,3]
To induce formation of Flk1+ vascular progenies, media containing Leukemia Inhibitory Factor (LIF) was removed from naive mouse embryonic stem (mES) and miPS cells, and the cells were cultured on IV collagen (IV Col)-coated dishes and propagated in presence of complete media containing BMP4, vascular endothelial growth factor (VEGF), and bFGF
We showed that miPS-derived cells gave rise to Flk1+ and Vascular Endothelial (VE)-cadherin+ endothelial cells (ECs) in a manner similar to ECs derived from mES cells
Summary
The transduction of fibroblast cells with transcription factors Nanog, Sox, Oct, Klf, and c-Myc converts these cells into induced pluripotent stem (iPS) cells [1,2,3]. The upstream components that induce exit of mesodermal cells to vascular cell progenies include factors such as bone morphogenetic proteins (BMPs), hypoxia, and PLOS ONE | www.plosone.org iPS-Derived Endothelial Cells. A major subset of mesodermal cells expressing Flk1+Flt1+VE-cadherin+CD34+CD31+ are capable of forming vascular plexus-like structures [20,21,22,23,24,25]. Flk1+ cells differentiated into ECs to form primitive vascular structures through the process of vasculogenesis [12,15,17,18,21]. Binding to vascular endothelial growth factor (VEGF) to Flk1/ VEGFR-2 regulates multiple aspects of neovascularization including EC development, survival, differentiation, migration, and lumenization [14,17,19,20,21]. The development of ECs entails timely expression and function of above key proteins
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