Development of astroglial cells in patterned neuronal cultures.

The purpose of this work was to study the development of astroglial cells in patterned neuronal cultures. Hippocampal neurons, derived from embryonic stage (E18) rats and cultured in serum-free Neurobasal/B27 medium, grew to follow patterns of poly(D-lysine) created by micro-contact printing. The growth of the astroglial cells and the co-localization of neurons and astroglial cells were measured for up to one month using fluorescence immunostaining of neurons and astroglial cells. Neurons grew to form square patterns within 2 weeks, while astroglia only started to emerge in the same period. Astroglial cells continued to proliferate for a month following a general growth curve. Over 90% of the astroglial cell area co-localized with neurons (within 2 mum) at an early stage of astroglial development (13 DIV). Over the remaining period, astroglial cells proliferated and the co-localization was 80%. Hence, in these culture conditions astroglial cells develop 2-3 weeks later than neurons but remain highly co-localized with neurons.