![]() In addition, we reported that Notch and β-catenin signaling are simultaneously activated in the downstream of cAMP and protein complex formation with Notch intracellular domain and β-catenin induced a set of arterial EC gene expressions resulted in arterial EC differentiation. In that same report, we also observed that PKA activation during ESC differentiation triggered EC differentiation and induced early commitment to EC lineage. We previously showed that PKA-activated CREB (cyclic AMP-responsive element (CRE) binding protein) bound to CRE on the Etv2/ER71 promoter region and directly induced Etv2/ER71 expression. Etv2/ER71, an ETS transcription factor, plays an indispensable role in EC and hematopoietic lineage commitment from early mesoderm. PKA is also directly involved in the EC commitment process. Coincidently, PKA activation increased the sensitivity of VEGFR2 + progenitors to VEGF, which increased the appearance of ECs also by a factor of ten. The binding of VEGF-A 165 to VEGFR2 and neuropilin1 is reported to enhance VEGFR signaling by approximately a factor of ten. Furthermore, we also found that cyclic adenosine monophosphate (cAMP) signaling potently enhances EC differentiation and that activation of a major downstream molecule of cAMP, protein kinase A (PKA), increased the expression of VEGFR2 and another VEGF receptor, neuropilin1, which together form a specific receptor for the VEGF-A 165 isoform. VEGF/VEGFR2 signaling is essential for inducing EC differentiation from VEGFR2-positive mesoderm cells. Previously, using mouse embryonic stem cells (ESCs), we established a method for systematic induction of cardiovascular cells from vascular endothelial growth factor (VEGF) receptor-2 (VEGFR2)-positive mesoderm cells as cardiovascular progenitors. Human pluripotent stem cells are one of the most suitable sources for such purpose. Thus, efficient EC preparation methods that provide scalable and stable supply are necessary for three-dimensional (3D) tissue engineering and organ regeneration. The importance of endothelial cells (ECs) has already been shown in the formation of various organs such as heart, liver, kidney, bone, and skin among many others. ![]() This does not alter our adherence to PLOS ONE policies on sharing data and materials.īlood vessels play essential roles in the generation of higher tissue structures, especially large tissue and organ structures. JKY, TI and HM are co-inventors on pluripotent stem cell-related patents. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the paper and its Supporting Information files.įunding: Support was provided by MEXT/JSPS KAKENHI Grant, #24390197 Core Center for iPS Cell Research, Research Center Network for Realization of Regenerative Medicine from Japan Agency for Medical Research and Development, AMED.Ĭompeting interests: JKY is a founder, equity holder, and scientific adviser of iHeart Japan Corporation. Received: OctoAccepted: FebruPublished: March 13, 2017Ĭopyright: © 2017 Ikuno et al. PLoS ONE 12(3):Įditor: Johnson Rajasingh, University of Kansas Medical Center, UNITED STATES (2017) Efficient and robust differentiation of endothelial cells from human induced pluripotent stem cells via lineage control with VEGF and cyclic AMP. We believe this method will be a valuable technological basis broadly for regenerative medicine and 3D tissue engineering.Ĭitation: Ikuno T, Masumoto H, Yamamizu K, Yoshioka M, Minakata K, Ikeda T, et al. This “stimulation-elimination” method robustly achieved very high efficiency (>99%) and yield (>10 ECs from 1 hiPSC input) of EC differentiation, with no purification of ECs after differentiation. We controlled the direction of differentiation from mesoderm to ECs using stage-specific stimulation with VEGF and cAMP combined with the elimination of non-responder cells at early EC stage. Here we report an efficient and robust EC differentiation method from human pluripotent stem cell lines based on a 2D monolayer, serum-free culture. We previously established an EC differentiation system with mouse pluripotent stem cells to show that vascular endothelial growth factor (VEGF) is essential to induce ECs and that cyclic adenosine monophosphate (cAMP) synergistically enhances VEGF effects. Thus, efficient induction of endothelial cells (ECs) from human pluripotent stem cells is a key method for generating higher tissue structures entirely from stem cells. Blood vessels are essential components for many tissues and organs.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |