High efficiency serum free feeder free erythroid differentiation of human pluripotent stem cells using small molecules

Olivier, E. N., Marenah, L., McCahill, A., Condie, A., Cowan, S. and Mountford, J. C. (2016) High efficiency serum free feeder free erythroid differentiation of human pluripotent stem cells using small molecules. Stem Cells Translational Medicine, 5(10), pp. 1394-1405. (doi:10.5966/sctm.2015-0371) (PMID:27400796) (PMCID:PMC5031182)

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Abstract

This article describes a good manufacturing practice (GMP)-compatible, feeder-free and serum-free method to produce large numbers of erythroid cells from human pluripotent stem cells (hPSCs), either embryonic or induced. This multistep protocol combines cytokines and small molecules to mimic and surpass the early stages of development. It produces, without any selection or sorting step, a population of cells in which 91.8% ± 5.4% express CD34 at day 7, 98.6% ± 1.3% express CD43 at day 10, and 99.1% ± 0.95% of cells are CD235a positive by day 31 of the differentiation process. Moreover, this differentiation protocol supports extensive expansion, with a single hPSC producing up to 150 hematopoietic progenitor cells by day 10 and 50,000–200,000 erythroid cells by day 31. The erythroid cells produced exhibit a definitive fetal hematopoietic type, with 90%–95% fetal globin and variable proportion of embryonic and adult globin at the protein level. The presence of small molecules during the differentiation protocol has quantitative and qualitative effects; it increases the proportion of adult globin and decreases the proportion of embryonic globin. Given its level of definition, this system provides a powerful tool for investigation of the mechanisms governing early hematopoiesis and erythropoiesis, including globin switching and enucleation. The early stages of the differentiation protocol could also serve as a starting point for the production of endothelial cells and other hematopoietic cells, or to investigate the production of long-term reconstituting hematopoietic stem cells from hPSCs.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:McCahill, Dr Angela and Marenah, Dr Lamin and Condie, Miss Alison and Cowan, Mr Scott and Mountford, Dr Joanne and Olivier, Dr Emmanuel
Authors: Olivier, E. N., Marenah, L., McCahill, A., Condie, A., Cowan, S., and Mountford, J. C.
College/School:College of Medical Veterinary and Life Sciences
College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
Journal Name:Stem Cells Translational Medicine
Publisher:AlphaMed Press
ISSN:2157-6564
ISSN (Online):2157-6580
Published Online:07 July 2016
Copyright Holders:Copyright © 2016 AlphaMed Press
First Published:First published in Stem Cells Translational Medicine 5(10):1394-1405
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
494631Proof of principle: Human embryonic stem cell derived red cell concentrates for clinical transfusionJoanne MountfordWellcome Trust (WELLCOME)087430/Z/08/ZRI CARDIOVASCULAR & MEDICAL SCIENCES