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Identification and conditions for selective expression of megakaryocytic
markers in Friend erythroleukemia cells
F Paoletti, AM Vannucchi, A Mocali, R Caporale and SA Burstein
Istituto di Patologia Generale, Universita' di Firenze, Italy.
Friend murine erythroleukemia cells (MELCs) have been reevaluated in terms
of their nature and potential pathways of differentiation. MELC induced
with 5 mmol/L hexamethylene bisacetamide (HMBA), in addition to expression
of known markers of the erythroid phenotype, were also found to exhibit
traits of the megakaryocytic lineage. Erythroid differentiation was shown
by the typical synthesis and accumulation of hemoglobin (Hb);
megakaryoblastoid differentiation of MELCs upon induction was shown by
increased specific activity of acetylcholinesterase (AChE). Incubation of
MELCs with 5 mmol/L HMBA in RPMI supplemented with 1% fetal calf serum
(FCS) (instead of the usual 5%), induced cells to selectively express high
levels of AChE (up to approximately 170 mU/mg protein) with little
activation of Hb synthesis (less than 5% B+ cells). The increase in AChE
levels was a general phenomenon affecting the whole cell population and
approached its maximum within 3 days of incubation with the inducer.
Subsequently, MELCs become committed to terminal division, undergoing
growth arrest and expression of the megakaryocytic phenotype even after the
removal of HMBA. There were no appreciable changes of basal AChE levels in
MELCs that were either made resistant to HMBA or treated with 0.1 mmol/L
hemin that activated differentiated erythroid function without commitment.
Phorbol 12-myristate 13-acetate (PMA), known to repress induced Hb
synthesis in these cells, did not prevent the full increase in AChE when
incubated with MELCs 2 days before HMBA addition. HMBA- induced MELCs
always underwent AChE increase that was more or less pronounced depending
on the low or high serum content in culture, respectively. Conversely, Hb
expression was permitted only when MELCs were transferred in the late phase
or at the end of commitment from low to high serum media. Variations of FCS
content in culture media proved to be a simple and reliable approach to
change the MELC response to inducers and to modulate expression of either
megakaryocytic or mixed erythromegakaryocytic phenotype. These findings
suggested that MELC might be considered, at least, as a bipotential model
of differentiation to be used for studies on regulation of either
megakaryocytic or erythroid markers and on competition between the two
hematopoietic lineages. In this regard, it was intriguing that AChE levels
attained under selective induction (low serum) were always higher than
under conditions allowing coexpression of both AChE and Hb (high serum).
Moreover, MELCs were also found to bind the specific rat- antimouse
platelet monoclonal antibody 4A5.(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 86,
Issue 7,
pp. 2624-2631,
10/01/1995
Copyright © 1995 by The American Society of Hematology
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