title: Characterization of mouse clonogenic megakaryocyte progenitors.
abstract: Although it has been shown that unfractionated bone marrow, hematopoietic stem cells, common myeloid progenitors, and bipotent megakaryocyteerythrocyte progenitors can give rise to megakaryocyte colonies in culture, monopotent megakaryocyte-committed progenitors (MKP) have never been prospectively isolated from the bone marrow of adult mice. Here, we use a monoclonal antibody to the megakaryocyte-associated surface protein, CD9, to purify MKPs from the c-kit(+)Sca-1(-)IL7Ralpha(-)Thy1.1(-)Lin(-) fraction of adult C57BLKa-Thy1.1 bone marrow. The CD9(+) fraction contained a subset of CD41(+)FcgammaR(lo)CD34(+)CD38(+) cells that represent approximately 0.01% of the total nucleated bone marrow cells. They give rise mainly to colony-forming unit-megakaryocytes and occasionally burst-forming unit-megakaryocytes, with a plating efficiency >60% at the single-cell level. In vivo, MKPs do not have spleen colony-forming activity nor do they contribute to long-term multilineage hematopoiesis; they give rise only to platelets for approximately 3 weeks. Common myeloid progenitors and megakaryocyteerythrocyte progenitors can differentiate into MKPs after 72 h in stromal cultures, indicating that MKPs are downstream of these two progenitors. These isolatable MKPs will be very useful for further studies of megakaryopoiesis as well as the elucidation of their gene expression patterns.
authors.affiliation: Department of Pathology and Developmental Biology, Stanford University School of Medicine, CA 94305, USA. nanakorn@leland.stanford.edu
authors.forename: Thanyaphong Na
authors.initials: TN
authors.lastname: Nakorn
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Miyamoto
authors.forename: Irving L
authors.initials: IL
authors.lastname: Weissman
mesh_terms: D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D004396:Coloring Agents; D016207:Cytokines / Q000494:pharmacology; D004247:DNA / Q000032:analysis; D008533:Megakaryocytes / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D013154:Spleen / Q000166:cytology; D013926:Thrombopoietin / Q000494:pharmacology
chemical_list: D000954:Antigens, Surface; D004396:Coloring Agents; D016207:Cytokines; D004247:DNA; D013926:Thrombopoietin