Adult human megakaryocyte-erythroid progenitors are in the CD34+CD38mid fraction.
Bipotent megakaryocyte/erythroid progenitors (MEPs) give rise to progeny limited to the megakaryocyte (Mk) and erythroid (E) lineages. We developed a novel dual-detection functional in vitro colony-forming unit (CFU) assay for single cells that differentiates down both the Mk and E lineages (CFU-Mk/E), which allowed development and validation of a novel purification strategy for the identification and quantitation of primary functional human MEPs from granulocyte colony-stimulating factor-mobilized peripheral blood and bone marrow. Applying this assay to fluorescence-activated cell sorter-sorted cell populations, we found that the Lin(-)CD34(+)CD38(mid)CD45RA(-)FLT3(-)MPL(+)CD36(-)CD41(-) population is much more highly enriched for bipotent MEPs than any previously reported subpopulations. We also developed purification strategies for primary human lineage-committed Mk and E progenitors identified as CFU-Mk and burst forming unit-E. Comparative expression analyses in MEP, MkP, and ErP populations revealed differential expression of MYB We tested whether alterations in MYB concentration affect the Mk-E fate decision at the single cell level in MEPs and found that short hairpin RNA-mediated MYB knockdown promoted commitment of MEPs to the Mk lineage, further defining its role in MEP lineage fate. There are numerous applications for these novel enrichment strategies, including facilitating mechanistic studies of MEP lineage commitment, improving approaches for in vitro expansion of Mk and E cells, and developing improved therapies for benign and malignant hematologic disease.
Department of Laboratory Medicine.
Chad
C
Sanada
Department of Laboratory Medicine, Yale Stem Cell Center.
Juliana
J
Xavier-Ferrucio
Department of Laboratory Medicine, Yale Stem Cell Center.
Yi-Chien
YC
Lu
Department of Cell Biology.
Elizabeth
E
Min
Department of Laboratory Medicine, Yale Stem Cell Center.
Ping-Xia
PX
Zhang
Yale Stem Cell Center, Department of Cell Biology.
Siying
S
Zou
Department of Laboratory Medicine.
Elaine
E
Kang
Yale Stem Cell Center, Department of Cell Biology.
Meng
M
Zhang
Department of Laboratory Medicine.
Gazelle
G
Zerafati
Yale Stem Cell Center, Department of Pediatrics, and.
Patrick G
0000-0002-5424-0224
PG
Gallagher
Department of Laboratory Medicine, Yale Stem Cell Center, Department of Cell Biology, Department of Pathology, Yale School of Medicine, New Haven, CT.
Diane S
0000-0001-9420-6947
DS
Krause
D051997:ADP-ribosyl Cyclase 1 / Q000378:metabolism*; D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism*; D019070:Cell Lineage; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D055015:Megakaryocyte-Erythroid Progenitor Cells / Q000166:cytology* / Q000378:metabolism; D008533:Megakaryocytes / Q000166:cytology; D010641:Phenotype; D020598:Proto-Oncogene Proteins c-myb / Q000378:metabolism; D053628:Receptors, Thrombopoietin / Q000378:metabolism; D051941:fms-Like Tyrosine Kinase 3 / Q000378:metabolism
D018952:Antigens, CD34; D020598:Proto-Oncogene Proteins c-myb; D053628:Receptors, Thrombopoietin; C082218:MPL protein, human; C497970:FLT3 protein, human; D051941:fms-Like Tyrosine Kinase 3; D051997:ADP-ribosyl Cyclase 1