Biopolym. Cell. 2022; 38(2):93-102.
Molecular and Cell Biotechnologies
Obtaining and characterization of rat fetuses neural cells aggregates/spheroids
1, 2Sukach O. M., 1Vsevolodska S. O., 1Ochenashko O. V., 1Shevchenko M. V.
  1. Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine
    23, Pereyaslavskaya Str., Kharkiv, Ukraine, 61015
  2. G. S. Skovoroda Kharkiv National Pedagogical University
    29, Alchevskyh Str., Kharkiv, Ukraine, 61002


Aim. Search for optimal conditions for the formation of aggregates/spheroids by neural cells (NC) isolated from the brain tissue of rat fetuses. Study of behavioral characteristics of NC in culture as a part of three-dimensional aggregates/spheroids. Methods. Isolation of NC from fetal rat brain tissue; cell viability assessment; cultivation; immunocytochemical staining; NC proliferative activity assessment. Results. It was found that the conditions for the robust formation of aggregates are: 1) the presence of 10 % adult rat’s blood serum in cell culture medium, 2) the seeding cell density – 1–4×106 cells/ml and 3) the initial cell viability higher than 20 %. Conclusions. The isolated rat fetuses brain cells, when cultured in the presence of 10 % serum and at concentrations above 1×106 cells/ml, spontaneously form multicellular aggregates. The structure of aggregates formed during short-term cultivation depends on the initial cell viability, and the size of aggregates depends on the initial viability and seeding densities of NC. The rat fetuses NC aggregates/spheroids contain stem/progenitor cells that can proliferate and differentiate. NC aggregates/spheroids form three-dimensional structures, in which favorable conditions are developed for the survival and adequate functioning of the committed and stem/progenitor NC.
Keywords: rat fetuses, neural cells, aggregates, spheroids, cultivation.


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