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

Emmanuel N. Olivier, Lamin Marenah, Angela McCahill, Alison Condie, Scott Cowan, Joanne C. Mountford

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1394-1405
Number of pages12
JournalStem cells translational medicine
Volume5
Issue number10
DOIs
StatePublished - Oct 1 2016
Externally publishedYes

Fingerprint

Pluripotent Stem Cells
Globins
Serum
Erythroid Cells
Hematopoietic Stem Cells
Stem Cells
Erythroid Precursor Cells
Erythropoiesis
Hematopoiesis
Embryonic Stem Cells
Endothelial Cells
Cytokines
Population
Proteins

Keywords

  • Cytokines
  • Erythropoiesis
  • Hematopoiesis
  • Pluripotent stem cells
  • Small molecule

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

High-efficiency serum-free feeder-free erythroid differentiation of human pluripotent stem cells using small molecules. / Olivier, Emmanuel N.; Marenah, Lamin; McCahill, Angela; Condie, Alison; Cowan, Scott; Mountford, Joanne C.

In: Stem cells translational medicine, Vol. 5, No. 10, 01.10.2016, p. 1394-1405.

Research output: Contribution to journalArticle

Olivier, Emmanuel N. ; Marenah, Lamin ; McCahill, Angela ; Condie, Alison ; Cowan, Scott ; Mountford, Joanne C. / High-efficiency serum-free feeder-free erythroid differentiation of human pluripotent stem cells using small molecules. In: Stem cells translational medicine. 2016 ; Vol. 5, No. 10. pp. 1394-1405.
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