TY - JOUR
T1 - High-efficiency serum-free feeder-free erythroid differentiation of human pluripotent stem cells using small molecules
AU - Olivier, Emmanuel N.
AU - Marenah, Lamin
AU - McCahill, Angela
AU - Condie, Alison
AU - Cowan, Scott
AU - Mountford, Joanne C.
N1 - Publisher Copyright:
© AlphaMed Press 2016.
PY - 2016/10/1
Y1 - 2016/10/1
N2 - This article describes a good manufacturing practice (GMP)-compatible, feeder-free and serum-free method to produce large numbers of erythroid cells from humanpluripotent 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.±%±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.
AB - This article describes a good manufacturing practice (GMP)-compatible, feeder-free and serum-free method to produce large numbers of erythroid cells from humanpluripotent 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.±%±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.
KW - Cytokines
KW - Erythropoiesis
KW - Hematopoiesis
KW - Pluripotent stem cells
KW - Small molecule
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U2 - 10.5966/sctm.2015-0371
DO - 10.5966/sctm.2015-0371
M3 - Article
C2 - 27400796
AN - SCOPUS:84988672601
SN - 2157-6564
VL - 5
SP - 1394
EP - 1405
JO - Stem Cells Translational Medicine
JF - Stem Cells Translational Medicine
IS - 10
ER -