The impact of fetal bovine serum on formation of spheroids from eye stromal limbal cells

Abstract

Widely used 3D culture technique, which allows obtaining cellular spheroids, opens up new opportunities for
modelling and studying pathological conditions, including ophthalmological ones, studying cellular characteristics
and behavior in environment close to in vivo conditions. Limbal zone is considered to be the main source of eye
stem cells for cornea regeneration under both normal and pathological conditions. Limbal multipotent
mesenchymal stromal cells that are the derivative of neural crest have a wide potential for differentiation and regeneration.
One of the up-to-date requirements for cell cultures, especially in clinical practice, is the use of serum-
free media. The current article is concerned with studying formation of cellular spheroids from primary
L-MMSC cell culture in growth media containing different concentrations of fetal bovine serum (FBS). We showed
that both in serum-free and serum-containing (1% and 10%) media cells formed viable spheroids by the 7th day. In
serum-free medium spheroids’ surface was performed with round cells, whereas addition of different concentrations
of serum resulted in formation of tegular surface layer. In the media containing serum the inner cells of
spheroids synthesized intercellular matrix, with the greater amount of matrix after culturing in medium with 10% of
FBS.