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J Michaeli, I Lerer, EA Rachmilewitz and E Fibach
The use of chromosome banding techniques has provided a valuable diagnostic
tool in various malignancies. The application of these methods, however, is
often restricted by a low yield of mitotic cells and the patient's
unwillingness to comply with repeated bone marrow aspiration. In an attempt
to promote mitotic activity of leukemic cells from the bone marrow and
peripheral blood, we employed a new method based on culturing the cells in
the presence of a conditioned medium derived from a human bladder carcinoma
cell line (5637). In addition to colony stimulating factor, this
conditioned medium contains a factor that is capable of stimulating
leukemic myeloblast proliferation. Bone marrow and peripheral blood
mononuclear cells from 58 patients with a variety of myeloid leukemias were
cultured for 24 to 120 hours in the presence or absence of conditioned
medium. These bone marrow cells showed a pronounced increase in the mitotic
index (5- to 50-fold) as compared to unstimulated cultures, and a greater
than 100-fold increase as compared to fresh, uncultured bone marrow cells.
Analyzable metaphases could be obtained even in marrow samples in which
direct or 24-hour G-banding techniques had failed to reveal metaphases. The
effect observed on peripheral blood cells was even more dramatic because
prior to culture no mitotic cells were detected, whereas up to 2% mitotic
cells were found in conditioned medium-stimulated peripheral leukemic
cells. Karyotype analysis of 36 out of the 58 leukemic patients has shown
that the chromosome changes discovered in conditioned medium stimulated
cells were identical to those found in unstimulated cells. New chromosome
aberrations, attributable to the stimulation of growth by conditioned
medium, were not found. The quality of the metaphases analyzed following
conditioned medium stimulation was considerably better than that of
unstimulated samples. Frozen cells, when cultured with conditioned medium,
were also suitable for cytogenetic analysis. Thus, the use of this
conditioned medium permits adequate cytogenetic analysis even in cases
where such analysis was previously impossible.
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| Copyright © 1986 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||