| Summary: | The in vitro biosynthesis of metallothionein (MT) has
been investigated in RBC precursors from human cord
blood in order to support the hypothesis for the
nucleated precursor origin of MT in human red blood
cells (RBC). Human RBC precursors are obtained by
(i) separating glycophorin A+ (gly A+) cells using a
magnetic cell sorting (MACS) technique and by (ii) ex
vivo expansion of precursors BFU-E (burst forming
unit-erythroid) on methylcellulose semi-solid culture
media from mononuclear cells of cord blood.
Biosynthesis of MT is detected at the protein level, by
immuno-histochemical staining using a mouse
monoclonal antibody (E9) in ex vivo expanded RBC
precursors obtained from BFU-E. Expression of MT is
also detected at the mRNA level by MT specific reverse
transcriptase polymerase chain reaction (RT-PCR)
both in ex vivo expanded precursors from BFU-E and
in MACS separated gly A+ cells. In addition, the expression of the fetal form of MT, MT-0 (also known
as MT-1H) at the mRNA level in glycophorin A+ cells,
is also confirmed by cDNA sequencing. With these
observations, to our knowledge, MT biosynthesis in
human erythroid precursors is reported for the first time.
Moreover, the current findings of MT-0 expression at
the mRNA level in gly A+ RBC precursors of hCB has
added one more member in the list of cells/organs like
fetal liver, human monocytes, non-neoplastic tissues
of adenocarcinoma etc., in which the expression of the
human fetal form of MT, i.e. MT-0, has also been
reported.
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