Isolation and characterization of human myeloid progenitor populations--TpoR as discriminator between common myeloid and megakaryocyte/erythroid progenitors

Experimental Hematology
Louise EdvardssonTor Olofsson

Abstract

The common myeloid progenitor (CMP) and its progeny, the granulocyte/monocyte progenitor (GMP) and megakaryocyte/erythrocyte progenitor (MEP), have been isolated based on the surface expression of CD34, IL-3Ralpha, and CD45RA. However, high resolution of IL-3Ralpha- and IL-3Ralphalo cells required to adequately separate the CMP and MEP populations is difficult to achieve. The aim of this study was to find a complementary surface marker to obtain a better separation of these two populations and to further characterize the acquired progenitor populations. To evaluate the thrombopoietin receptor (TpoR) as a candidate marker, CD19-/CD34+/IL-3Ralphalo/-/CD45RA-/TpoR- (CMP), CD19-/CD34+/IL-3Ralphalo/CD45RA+/TpoR- (GMP), and CD19-/CD34+/IL-3Ralphalo/-/CD45RA-/TpoR+ (MEP) cells from human bone marrow were sorted to semisolid cultures for colony assays, and in addition analyzed for their surface expression of other growth factor receptors (GFRs) and sorted to real-time RT-PCR for gene expression analysis. The colony-formation and gene expression assays showed that inclusion of TpoR as a marker gave a distinct and reproducible separation of the myeloid progenitors. Furthermore, most GFR surface expression correlated to gene expression, b...Continue Reading

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