THE OPTIMIZATION METHOD OF ISOLATION OF IMMATURE DENDRITIC CELLS
A.N. Goltsev, T.G. Dubrava, E.E. Yampolskaya, Yu.A. Gayevskaya, N.N. Babenko, N.A. Bondarovich, I.G. Kovalenko
Institute for Problems of Cryobiology and Cryomedicine
of the National Academy of Sciences of Ukraine, Kharkiv,
The method of isolation of bone marrow mononuclear cells
in «trasograph» gradient, enabling to obtain more than 40
% of cells, among those 28 % were CD14+-monocytes has
been presented. The minimum set of phenotype markers to
assess immature dendritic cells in vitro has been selected. In
a comparative aspect it has been shown that the most adequate
sources of obtaining of dendritic cells are mononuclear cells in
contrast to total cell suspension of bone marrow. It has been
found that the differentiation of myeloid progenitor cells and
monocytes into immature dendritic cells was accompanied
with the loss of CD14 antigen , minimization of expression
rate of co-stimulatory molecule CD86 and appearance of
CD11b-antigen marker, which is the feature of these cells. The
results allow not only to estimate the qualitative characteristics
of the obtained dendritic cells and their tolerogenic activity in
the model of autoimmune diseases, but also to underline the
perspectives of establishing their stocks at low temperature
banks to be used in clinics.
dendritic cells; bone marrow; mononuclear cells, monocytes; co-stimulatory molecules
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