Scalable Expansion of CD71+ Erythroid Progenitor Cells for Cell Therapy
AssigneeUniversity of Pittsburgh—Of the Commonwealth System of Higher Education
InventorsMo Reza Ebrahimkhani, Ryan LeGraw
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Abstract
Provided herein is a scalable method of producing CD71+ erythroid progenitor cells for use in cell therapies, such as immunomodulatory therapies in which a T-cell response is suppressed. The method includes induction of a fetal liver/yolk sac organoid from pluripotent stem cells. e.g. induced pluripotent stem cells by GATA6, followed by co-culturing CD34+ cells. e.g. from cord blood or bone marrow, with the organoid, optionally in the presence of thrombopoietin, stem cell factor, and/or FLT3 ligand. Also provided herein is a bioreactor for producing the CD71+ erythroid progenitor cells.
Claims27
A method of preparing a population of CD71+ cells, comprising:
- culturing pluripotent stem cells to differentiate to a fetal liver/yolk sac phenotype, such as increased expression of erythroid markers CD36, CD47, erythropoietin receptor (EPOR), and transforming growth factor β1 (TGFβ1);
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- coculturing CD34+ hematopoietic stem and progenitor cells (HPSCs) with the differentiated pluripotent stem cells to produce an expanded population of CD71+ (CD71HI) cells from the HPSCs.
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A cell culture device comprising:
- a cell culture vessel comprising cell culture media, and a coculture of cells having a fetal liver/yolk sac phenotype and CD34+ hematopoietic stem and progenitor cells.
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Dependencies are inferred from the claim text.