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Blood, 15 September 2006, Vol. 108, No. 6, pp. 2095-2105.
Prepublished online as a Blood First Edition Paper on June 6, 2006; DOI 10.1182/blood-2006-02-003327.
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Submitted February 10, 2006
Accepted May 13, 2006
Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures
Maxim A Vodyanik, James A Thomson, and Igor I Slukvin*
National Primate Research Center, University of Wisconsin Graduate School, Madison, WI, USA
* Corresponding author; email: islukvin{at}wisc.edu.
During hematopoietic differentiation of human embryonic stem cells (hESC), early hematopoietic progenitors arise along with endothelial cells within CD34+ population. Although hESC-derived hematopoietic progenitors have been previously identified by functional assays, their phenotype has not been defined. Here, using hESC differentiation in coculture with OP9 stromal cells, we demonstrated that early progenitors committed to hematopoietic development could be identified by surface expression of leukosialin (CD43). CD43 was detected on all types of emerging clonogenic progenitors before expression of CD45, persisted on differentiating hematopoietic cells, and reliably separated hematopoietic CD34+ population from CD34+CD43-CD31+KDR+ endothelial and CD34+CD43-CD31-KDR- mesenchymal cells. Furthermore, we demonstrated that the first-appearing CD34+CD43+CD235a+CD41a+/-CD45- cells represent pre-committed erythro-megakaryocytic progenitors. Multipotent lymphohematopoietic progenitors were generated later as CD34+CD43+CD41a-CD235a-CD45- cells. These cells were negative for lineage-specific markers (Lin-), expressed KDR, VE-cadherin, and CD105 endothelial proteins, and GATA-2, GATA-3, RUNX1, c-myb transcription factors that typify initial stages of definitive hematopoiesis originating from endothelial-like precursors. Acquisition of CD45 expression by CD34+CD43+CD45-Lin- cells was associated with progressive myeloid commitment and a decrease of B-lymphoid potential. CD34+CD43+CD45+Lin- cells were largely devoid of VE-cadherin and KDR expression, and had a distinct Flt-3highGATA-3lowRUNX1lowPU1highMPOhighIL7R high gene expression profile.
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