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In vitro establishment and characterization of a human megakaryoblastic
cell line
DA Fugman, DP Witte, CL Jones, BJ Aronow and MA Lieberman
Department of Hematology-Oncology, Children's Hospital Medical Center,
Cincinnati, OH.
A human megakaryoblastic cell line, designated CHRF-288-11, has been
established in vitro through the use of adherent stromal cells in long-
term human bone marrow culture. Long-term bone marrow cultures were
required for the initial adaptation of the megakaryoblastic cells to
culture conditions; however, once adapted, the cells were weaned from the
stromal layer until they proliferated in the complete absence of any feeder
layers. The seed cells for the establishment of this line were derived from
a solid tumor; the cloned cell line derived from this tumor exhibits
markers characteristic of megakaryocytes and platelets. Specifically, the
cells express platelet peroxidase, platelet factor 4, and platelet
Ca+(+)-adenosine triphosphatase (ATPase), glycoprotein IIb- IIIa (CDw41),
factor VIII antigen, and the MY7 (CD13) and MY9 (CD33) antigens. The cells
do not express the erythroid markers glycophorin A and hemoglobin, the
myeloid marker myeloperoxidase, nor markers specific for T and/or B cells.
The established cell line produces both basic fibroblast growth factor and
transforming growth factor-beta, properties demonstrated previously for the
solid tumor. The clonal cell population exhibited a unique, singular
karyotype, indicating cellular homogeneity. The cells display a doubling
time of approximately 33 hours in either 25% horse or calf serum. Treatment
of the cells with 1 X 10(-8) mol/L phorbol 12-myristate 13-acetate (PMA)
leads to the induction of multi-nucleation and hyperploidy in the cells,
with approximately 35% of the cells exhibiting two or more nuclei per cell,
and greater than 80% of the cells enlarging in size. The establishment of
this unique cell line under defined culture conditions will be beneficial
for the future study of megakaryocytic properties expressed by this cell
line.
Volume 75,
Issue 6,
pp. 1252-1261,
03/15/1990
Copyright © 1990 by The American Society of Hematology
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