Obtaining and characterization of rat fetuses neural cells aggregates/spheroids

Abstract

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 the cell culture medium, 2) the seeding cell density -1-4×10 6 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 1×10 6 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.