Cargando…

Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells

Embryonic stem cells (ESCs) could potentially compensate for the lack of blood platelets available for use in transfusions. Here, we describe a new method for generating mouse ESC-derived platelets (ESPs) that can contribute to hemostasis in vivo. Flow cytometric sorting of cells from embryoid bodie...

Descripción completa

Detalles Bibliográficos
Autores principales: Nishikii, Hidekazu, Eto, Koji, Tamura, Noriko, Hattori, Koichi, Heissig, Beate, Kanaji, Taisuke, Sawaguchi, Akira, Goto, Shinya, Ware, Jerry, Nakauchi, Hiromitsu
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525582/
https://www.ncbi.nlm.nih.gov/pubmed/18663123
http://dx.doi.org/10.1084/jem.20071482
_version_ 1782158667796185088
author Nishikii, Hidekazu
Eto, Koji
Tamura, Noriko
Hattori, Koichi
Heissig, Beate
Kanaji, Taisuke
Sawaguchi, Akira
Goto, Shinya
Ware, Jerry
Nakauchi, Hiromitsu
author_facet Nishikii, Hidekazu
Eto, Koji
Tamura, Noriko
Hattori, Koichi
Heissig, Beate
Kanaji, Taisuke
Sawaguchi, Akira
Goto, Shinya
Ware, Jerry
Nakauchi, Hiromitsu
author_sort Nishikii, Hidekazu
collection PubMed
description Embryonic stem cells (ESCs) could potentially compensate for the lack of blood platelets available for use in transfusions. Here, we describe a new method for generating mouse ESC-derived platelets (ESPs) that can contribute to hemostasis in vivo. Flow cytometric sorting of cells from embryoid bodies on day 6 demonstrated that c-Kit(+) integrin αIIb (αIIb)(+) cells, but not CD31(+) cells or vascular endothelial cadherin(+) cells, are capable of megakaryopoiesis and the release of platelet-like structures by day 12. αIIbβ3-expressing ESPs exhibited ectodomain shedding of glycoprotein (GP)Ibα, GPV, and GPVI, but not αIIbβ3 or GPIbβ. ESPs showed impaired αIIbβ3 activation and integrin-mediated actin reorganization, critical events for normal platelet function. However, the administration of metalloproteinase inhibitors GM6001 or TAPI-1 during differentiation increased the expression of GPIbα, improving both thrombogenesis in vitro and posttransfusion recovery in vivo. Thus, the regulation of metalloproteinases in culture could be useful for obtaining high-quality, efficacious ESPs as an alternative platelet source for transfusions.
format Text
id pubmed-2525582
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-25255822009-02-04 Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells Nishikii, Hidekazu Eto, Koji Tamura, Noriko Hattori, Koichi Heissig, Beate Kanaji, Taisuke Sawaguchi, Akira Goto, Shinya Ware, Jerry Nakauchi, Hiromitsu J Exp Med Articles Embryonic stem cells (ESCs) could potentially compensate for the lack of blood platelets available for use in transfusions. Here, we describe a new method for generating mouse ESC-derived platelets (ESPs) that can contribute to hemostasis in vivo. Flow cytometric sorting of cells from embryoid bodies on day 6 demonstrated that c-Kit(+) integrin αIIb (αIIb)(+) cells, but not CD31(+) cells or vascular endothelial cadherin(+) cells, are capable of megakaryopoiesis and the release of platelet-like structures by day 12. αIIbβ3-expressing ESPs exhibited ectodomain shedding of glycoprotein (GP)Ibα, GPV, and GPVI, but not αIIbβ3 or GPIbβ. ESPs showed impaired αIIbβ3 activation and integrin-mediated actin reorganization, critical events for normal platelet function. However, the administration of metalloproteinase inhibitors GM6001 or TAPI-1 during differentiation increased the expression of GPIbα, improving both thrombogenesis in vitro and posttransfusion recovery in vivo. Thus, the regulation of metalloproteinases in culture could be useful for obtaining high-quality, efficacious ESPs as an alternative platelet source for transfusions. The Rockefeller University Press 2008-08-04 /pmc/articles/PMC2525582/ /pubmed/18663123 http://dx.doi.org/10.1084/jem.20071482 Text en © 2008 Nishikii et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jem.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Articles
Nishikii, Hidekazu
Eto, Koji
Tamura, Noriko
Hattori, Koichi
Heissig, Beate
Kanaji, Taisuke
Sawaguchi, Akira
Goto, Shinya
Ware, Jerry
Nakauchi, Hiromitsu
Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title_full Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title_fullStr Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title_full_unstemmed Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title_short Metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
title_sort metalloproteinase regulation improves in vitro generation of efficacious platelets from mouse embryonic stem cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2525582/
https://www.ncbi.nlm.nih.gov/pubmed/18663123
http://dx.doi.org/10.1084/jem.20071482
work_keys_str_mv AT nishikiihidekazu metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT etokoji metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT tamuranoriko metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT hattorikoichi metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT heissigbeate metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT kanajitaisuke metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT sawaguchiakira metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT gotoshinya metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT warejerry metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells
AT nakauchihiromitsu metalloproteinaseregulationimprovesinvitrogenerationofefficaciousplateletsfrommouseembryonicstemcells