Growth and differentiation of human stem cell factor/erythropoietin-dependent erythroid progenitor cells in vitro.
Stem cell factor (SCF) and erythropoietin (Epo) effectively support erythroid cell development in vivo and in vitro. We have studied here an SCF/Epo-dependent erythroid progenitor cell from cord blood that can be efficiently amplified in liquid culture to large cell numbers in the presence of SCF, Epo, insulin-like growth factor-1 (IGF-1), dexamethasone, and estrogen. Additionally, by changing the culture conditions and by administration of Epo plus insulin, such progenitor cells effectively undergo terminal differentiation in culture and thereby faithfully recapitulate erythroid cell differentiation in vitro. This SCF/Epo-dependent erythroid progenitor is also present in CD34(+) peripheral blood stem cells and human bone marrow and can be isolated, amplified, and differentiated in vitro under the same conditions. Thus, highly homogenous populations of SCF/Epo-dependent erythroid progenitors can be obtained in large cell numbers that are most suitable for further biochemical and molecular studies. We demonstrate that such cells express the recently identified adapter protein p62(dok) that is involved in signaling downstream of the c-kit/SCF receptor. Additionally, cells express the cyclin-dependent kinase (CDK) inhibitors p21(cip1) and p27(kip1) that are highly induced when cells differentiate. Thus, the in vitro system described allows the study of molecules and signaling pathways involved in proliferation or differentiation of human erythroid cells.
Panzenböck
B
B
Max-Delbrück-Centre for Molecular Medicine, MDC, Berlin, Germany; and the Humboldt University Berlin, Virchow Klinikum, Robert-Rössle-Klinik, Berlin, Germany.
Bartunek
P
P
Mapara
M Y
MY
Zenke
M
M
D001773:Blood Cells / Q000166:cytology / Q000187:drug effects; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D018797:Cell Cycle Proteins; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D050759:Cyclin-Dependent Kinase Inhibitor p21; D050760:Cyclin-Dependent Kinase Inhibitor p27; D016213:Cyclins / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins; D003907:Dexamethasone / Q000494:pharmacology; D004790:Enzyme Induction; D015672:Erythroid Precursor Cells / Q000166:cytology* / Q000187:drug effects; D004920:Erythropoiesis / Q000187:drug effects*; D004921:Erythropoietin / Q000494:pharmacology; D004967:Estrogens / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics; D009928:Organ Specificity; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics; D016601:RNA-Binding Proteins; D015398:Signal Transduction; D019089:Stem Cell Factor / Q000494:pharmacology; D025521:Tumor Suppressor Proteins
C493956:CDKN1A protein, human; D018797:Cell Cycle Proteins; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D004268:DNA-Binding Proteins; C489560:DOK1 protein, human; D004967:Estrogens; C074561:GAP-associated protein p62; D007328:Insulin; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D019089:Stem Cell Factor; D025521:Tumor Suppressor Proteins; D004921:Erythropoietin; D050760:Cyclin-Dependent Kinase Inhibitor p27; D007334:Insulin-Like Growth Factor I; D003907:Dexamethasone