Transduction of recombinant human erythropoietin receptor cDNA into daughter progenitors derived from single CD34(3+) cord blood cells changes the differentiation profile of daughter progenitors.
In this study, we tested the capacity to change the differentiation profile of progenitor cells by retroviral-mediated transduction of EpoR cDNA into one of the paired daughter cells derived from single CD34(3+) CB cells. Our results show that for the non-viral-treated daughter cells, the majority (99.6%) formed the same colony type. However, with cells transduced with viral vectors, 7.1% of the daughter cells transduced with the EpoR cDNA formed either a burst forming unit-erythroid (BFU-E) or a colony-forming unit-granulocyte, macrophage, erythroid, megakaryocyte (CFU-GEMM) colony compared to the other daughter cell transduced with viral supernatant lacking EpoR cDNA, which formed either a colony-forming unit granulocyte-macrophage (CFU-GM) or a high proliferative potential-colony forming cell (HPP-CFC) colony. Expression of the transduced EpoR cDNA was confirmed in individual colonies by RT-PCR analysis. These results substantiate in a more rigorous fashion our previous results that it is possible to change the Epo-responsive differentiation profile of progenitor cells by transduction into these cells of an EpoR cDNA and this change was apparent only in daughter cells derived from single CD34(3+) kit+ cells transduced with EpoR cDNA.
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Department of Microbiology, Walther Oncology Center, Indiana University School of Medicine, Indianapolis 46202-5121, USA.
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Rice
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Broxmeyer
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D015703:Antigens, CD / Q000502:physiology; D018952:Antigens, CD34 / Q000502:physiology*; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D018076:DNA, Complementary; D005312:Fetal Blood; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology / Q000502:physiology*; D006801:Humans; D007231:Infant, Newborn; D016133:Polymerase Chain Reaction; D017467:Receptors, Erythropoietin / Q000096:biosynthesis / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012190:Retroviridae; D014162:Transfection
D015703:Antigens, CD; D018952:Antigens, CD34; D018076:DNA, Complementary; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins