Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells

Kyung Dal Choi, Maxim Vodyanik, Igor I. Slukvin

Research output: Contribution to journalArticlepeer-review

93 Scopus citations

Abstract

In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin ĝ̂' CD34 + CD43 + CD45 + multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5g-19 d of culture with cytokines, depending on the cell type.

Original languageEnglish (US)
Pages (from-to)296-313
Number of pages18
JournalNature Protocols
Volume6
Issue number3
DOIs
StatePublished - Mar 2011

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