Cord blood megakaryocytes do not complete maturation, as indicated by impaired establishment of endomitosis and low expression of G1/S cyclins upon thrombopoietin-induced differentiation.
Cord blood (CB) has successfully been used as a stem cell source for haemopoietic reconstitution. However, a significant delay in platelet engraftment is consistently found in CB versus adult peripheral blood (PB) or bone marrow transplants. We sought to determine whether or not CB megakaryocytes have reached terminal maturation and, hence, full thrombopoietic potential. A comparative analysis of megakaryocytes cultured from either CB or PB progenitors in the presence of thrombopoietin (TPO) showed a similar differentiation response, although proliferation was 2.4 times higher in CB than in PB cells. Importantly, the TPO-induced ploidy level was notably different: whereas 82.7% of CB megakaryocytes remained diploid (2N) at the end of the culture, more than 50% of PB megakaryocytes had reached a DNA content equal to or higher than 4N. Western blot and flow cytometry analyses revealed that only polyploid PB megakaryocytes expressed cyclins E, A and B, whereas cyclin D3 was detected in both fetal and adult megakaryocytic nuclei. These data suggest that establishment of endomitotic cycles is impaired in CB megakaryocytes, associated with a differential regulation of G1/S cell cycle factors. We believe that the relative immaturity of fetal megakaryocytes could be a contributing factor to the delayed platelet engraftment in cord blood transplantation.
Bornstein
R
R
Madrid Cord Blood Bank, Hospital Universitario 12 de Octubre, Madrid, Spain.
García-Vela
J
J
Gilsanz
F
F
Auray
C
C
Calés
C
C
D015703:Antigens, CD / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D016213:Cyclins / Q000378:metabolism*; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D039661:Integrin beta3; D008533:Megakaryocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000276:immunology; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D013926:Thrombopoietin / Q000494:pharmacology*; D013997:Time Factors
D015703:Antigens, CD; D018952:Antigens, CD34; D016213:Cyclins; D039661:Integrin beta3; D010980:Platelet Membrane Glycoproteins; D013926:Thrombopoietin