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Blood, 1 February 2002, Vol. 99, No. 3, pp. 888-897
HEMATOPOIESIS
Different ploidy levels of megakaryocytes generated from
peripheral or cord blood CD34+ cells are correlated with
different levels of platelet release
Gianfranco Mattia,
Francesca Vulcano,
Luisa Milazzo,
Alessandra Barca,
Giampiero Macioce,
Adele Giampaolo, and
H. Jane Hassan
From the Department of Clinical Biochemistry, Istituto
Superiore di Sanità, Rome, Italy.
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
(~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 (~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.
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