Influence of lineage-specific cytokines on commitment and asymmetric cell division of haematopoietic progenitor cells.
We examined the influence of cytokines on erythroid- and myeloid-lineage development of AC133+ cells during primary and secondary cultures. Cells cultured for 14 d in liquid medium containing erythropoietin (EPO) were amplified 831-fold with 98.2% erythroid cells. A similar culture exposed to granulocyte colony-stimulating factor (G-CSF) grew 1350-fold with 97.4% myeloid cells. To assess whether the cells with EPO inducement could respond at this point to G-CSF signal, or vice versa, the EPO-stimulated population was re-grown with G-CSF, constituting 95.2% myeloid, of 5075-fold, cells after 14 d of re-culture. Conversely, reculture of the G-CSF-stimulated population with EPO resulted in a 4083-fold growth with 81.4% erythroid cells. Semisolid culture containing EPO orG-CSF showed that some individual colonies had self- renewal potential after 14 d culture and could be induced todevelop into a different lineage. Analysis of primitive markers, CD34 and Notch1, or lineage markers, EPO-R and CD13, by single-cell reverse transcription polymerase chain reaction showed that individual colonies of 2-16 cells contained at least one CD34-positive cell with expression ofNotch1 and co-expression of EPO-R and CD13 appeared on either CD34-positive or CD34-negative cells. In situ hybridization with the same cell surface markers in cell populations confirmed the asymmetric cell division and co-expression from single cell data. The study provides a useful model for the analysis of multipotential progenitor development, and indicates that progenitor cells co-express genes from different lineage pathways before commitment and that cytokines influence lineage commitment.
Chen
Ling
L
Molecular and Clinical Hematology Branch, National Institute of Diabetes and Digestive and Kidney Disease, National Institutes of Health, Bethesda, MD 20892, USA.
Zhang
Jiachang
J
Tang
Delia C
DC
Fibach
Eitan
E
Rodgers
Griffin P
GP
D000071916:AC133 Antigen; D015703:Antigens, CD; D000954:Antigens, Surface / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003114:Colony-Forming Units Assay; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000187:drug effects; D004921:Erythropoietin / Q000494:pharmacology*; D006023:Glycoproteins / Q000378:metabolism; D016179:Granulocyte Colony-Stimulating Factor / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000187:drug effects; D010455:Peptides / Q000378:metabolism
D000071916:AC133 Antigen; D015703:Antigens, CD; D000954:Antigens, Surface; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D004921:Erythropoietin; D016179:Granulocyte Colony-Stimulating Factor