Obtaining a three-cell model of a neurovascular unit in vitro

Abstract

At present, there are many ways to model the blood–brain barrier and neurovascular unit in vitro. All the proposed models have advantages and disadvantages related to the specifics of their obtaining and use. This work is dedicated to a study of possibility to obtain an in vitro three-cell model of a neurovascular unit (NVU) based on progenitor cells isolated from brain of rat embryos. From the obtained progenitor cells, we cultured neurospheres with subsequent differentiation into astrocytes and neurons. In addition, from embryonic cells, we isolated cell endotheliocytes. The direct object of observation was cell cocultures in Petri dishes. In differentiation of progenitor calls, the formation of a monolayer of astrocytes occurs in 7–9 days; for neurons, in 10–14 days; and for endotheliocytes, in 7 days. The developed protocol of simultaneous isolation and cultivation of all cell types makes it possible to shorten periods of obtaining three-cell NVU model in vitro and reduce the number of used animals, as well as the number of supplies and reagents.