MORPHOFUNCTIONAL FEATURES OF FIBROBLASTS LINE L929 IN 3D-CULTURE
G.A. Bozhok, A.I. Moisieiev, O.L. Gorina, Т.P. Bondarenko
Institute for Problems of Cryobiology and Cryomedicine of the NAS of Ukraine, Kharkiv, Ukraine
DOI: https://doi.org/10.15407/fz65.03.034
Abstract
The aim of the work was to standardize the conditions for
obtaining viable and functionally complete cells in the
composition of multicellular spheroids (MS). The latter were
obtained in the conditions of 7 days 3D cultivation with fibroblasts
seeding at concentrations of 1·105, 2·105 and 5·105 cells/
ml. It has been proved that the sowing concentration of fibroblasts
affects the size of the formed MS as well as the viability,
morphological parameters, proliferative and adhesive properties
of cells they are composed of. The maximal concentration that
have been used had a negative impact on spheroid formation,
on the ability of cells to adhesion. This concentration reduced
cell viability by 19%, coefficient of increasing the number
of cells 4,25-fold and increased the number of cells with
nuclear fragmentation and chromatin condensation by 20% in
comparison with the sowing concentration of 2·105 cells/ml.
At a concentration of 1·105 cells/ml, predominantly single and
small (about 10-20 mkm) MS with a cell viability of about 89,5 ±
1,2% were formed. The fibroblasts had normal morphological
characteristics, but the coefficient of increasing the number
of cells was significantly lower than the similar indicator at
the sowing concentration of 2·105 cells/ml. Thus, it has been
established that the concentration of 2·105 cells/ml is optimal
for the formation of MS with morphologically and functionally
complete fibroblasts.
Keywords:
fibroblasts of L929 cell line; 3D-cultivation; multicellular spheroids; viability, proliferative and adhesive properties of cells in the composition of multicellular spheroids.
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