RRC ID 4343
Author Takayama N, Nishikii H, Usui J, Tsukui H, Sawaguchi A, Hiroyama T, Eto K, Nakauchi H.
Title Generation of functional platelets from human embryonic stem cells in vitro via ES-sacs, VEGF-promoted structures that concentrate hematopoietic progenitors.
Journal Blood
Abstract Human embryonic stem cells (hESCs) could potentially represent an alternative source for blood transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. When we cultured hESCs on either C3H10T1/2 or OP-9 cells to facilitate hematopoiesis, we found that exogenous administration of vascular endothelial growth factor promoted the emergence of sac-like structures, which we named embryonic stem cell-derived sacs (ES-sacs). These ES-sacs consisted of multiple cysts demarcated by cellular monolayers that retained some of the properties of endothelial cells. The spherical cells inside ES-sacs expressed primarily CD34, along with VE-cadherin, CD31, CD41a, and CD45, and were able to form hematopoietic colonies in semisolid culture and to differentiate into mature megakaryocytes by day 24 in the presence of thrombopoietin. Apparently, ES-sacs provide a suitable environment for hematopoietic progenitors. Relatively large numbers of mature megakaryocytes could be induced from the hematopoietic progenitors within ES-sacs, which were then able to release platelets that displayed integrin alpha IIb beta 3 activation and spreading in response to ADP or thrombin. This novel protocol thus provides a means of generating platelets from hESCs, which could serve as the basis for efficient production of platelets for clinical transfusion and studies of thrombopoiesis.
Volume 111(11)
Pages 5298-306
Published 2008-6-1
DOI 10.1182/blood-2007-10-117622
PII blood-2007-10-117622
PMID 18388179
MeSH Blood Platelets / cytology* Blood Platelets / metabolism Cell Culture Techniques / methods* Embryonic Stem Cells / cytology* Embryonic Stem Cells / metabolism Flow Cytometry Hematopoietic Stem Cells / cytology* Hematopoietic Stem Cells / metabolism Humans Immunohistochemistry Megakaryocytes / cytology Megakaryocytes / metabolism Microscopy, Confocal Platelet Activation / physiology Reverse Transcriptase Polymerase Chain Reaction Vascular Endothelial Growth Factor A / metabolism*
IF 15.132
Times Cited 141
Human embryonic stem cells KhES-1 KhES-2 KhES-3