A New Method in Cns (Central Nervous System) in Vitro Cultures in the Mouse: Study of Effectiveness

Abstract

In this study: to evaluate the effectiveness of collagen coating method, using in the peripheral nervous system cultures. and its involving factors caused from manipulations in central nervous system (CNS) cultures was aimed. Via frontal approach, brains, transected from young Swiss albino mice, were taken into artificial cerebro-spinal fluid immediately and made blocks in agarose gel. With a vibration microtome, 200 pm thickness horizontally live slices were taken in to the dishes filled with culture medium. Tissue sections were analyzed as two groups. In the group 1 (control): fresh slices were evaluated directly. In the group 2: sections were covered with collagen gel (Type I) and left in the incubator (5% CO(2)) for 3 days. These sections were dyed with calcein and propidium iodide for viability and non-viability and then observed with confocal laser scanning microscope. Images were captured digitally and examined. Since negative effects of high melting temperature of standard agar on the livability, using low melting agar to tissue blocking and high frequency - low speed vibrotome setting to cut were more preferably. In the 3 days cultures, viability/nonviability rates were indicated better values. It is concluded that, in the CNS slicing cultures, collagen coating method was an easier, effective, useful and alternative method to present techniques.