Different ploidy levels of megakaryocytes generated from peripheral or cord blood CD34+ cells are correlated with different levels of platelet release.
Ploidy could be the key to understanding megakaryocyte (MK) biology and platelet production. Human CD34+ cells purified from umbilical cord blood (CB) and peripheral blood (PB) were investigated on their capability to give rise, in a serum-free medium containing thrombopoietin, to MKs and platelets. CB-MKs showed reduced polyploidization and platelet number compared with PB-MKs, but a similar membrane phenotype. Most CB-MKs showed a 2N content of DNA (approximately 80%) and only 2.6% had 8N, whereas 40% of the PB cells had 8N or more. Platelets were substantially released in PB culture from day 12; at day 14 the CB-derived MKs were able to release platelets although at a reduced level (approximately 35%), correlating with their reduced size. A direct correlation was demonstrated by sorting polyploid cells from PB-MKs and evaluating the platelets released in the supernatant. Furthermore, the study analyzed the expression and distribution of cyclin D3 and cyclin B1. Cyclin D3 protein was increased in PB in comparison to CB-MKs; in PB culture most cells rapidly became positive, whereas in CB-derived cells cyclin D3 expression was evident only from day 9 and in a reduced percentage. Cyclin B1 was essentially localized at the nuclear level in the CB and was expressed during the whole culture. In PB-MKs, at day 9, a reduction was observed, correlating with an advanced ploidy state. The data indicate the inability of the CB-MKs to progress in the endomitotic process and a direct correlation between DNA content and platelet production.
Mattia
Gianfranco
G
Department of Clinical Biochemistry, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
Vulcano
Francesca
F
Milazzo
Luisa
L
Barca
Alessandra
A
Macioce
Giampiero
G
Giampaolo
Adele
A
Hassan
H Jane
HJ
D000328:Adult; D018952:Antigens, CD34 / Q000032:analysis*; D001772:Blood Cell Count; D001773:Blood Cells / Q000166:cytology / Q000276:immunology*; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism*; D002454:Cell Differentiation / Q000187:drug effects; D018925:Chemokines / Q000032:analysis / Q000502:physiology; D016213:Cyclins / Q000032:analysis / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D016130:Immunophenotyping; D007231:Infant, Newborn; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D011003:Ploidies; D013926:Thrombopoietin / Q000494:pharmacology
D018952:Antigens, CD34; D018925:Chemokines; D016213:Cyclins; D013926:Thrombopoietin