Attachment and Culture of Dissociated Cells From Rat Embryo Cerebral Hemispheres on Polylysine-Coated Surface

Abstract

INTRODUCTIONhemispheres from 16 to 18-day old rat embryo were treated with a solution of 0 .125% (wt/vol) trypsin for 5 min, and the resulting cell suspension was aspirated through a Pasteur pipette to complete the dissociation . The cells were centrifuged at 400 g for 5 min and the supernate was discarded . The pellet was then resuspended in the growth medium (13), and the suspension was centrifuged for I min at 200 g . The supernate containing the dissociated single cells was transferred to a sterile bottle, final cell concentration was adjusted to 0 .1 0 .3 x 106 cells per I ml medium, and 3 ml of the above suspension was placed into polylysine-coated Petri dishes . After 30 min incubation at 37üC in an atmosphere of 95% air, 5% CO 2 , the medium containing the nonadhered cells was removed and replaced by fresh medium which was then changed every 3-4 days, depending on its acidity . Phase-contrast microscopy was employed for observation of cultures during their growth period .

Distinct Properties of Neuronal and Astrocytic Endopeptidase 3.4.24.16: A Study on Differentiation, Subcellular Distribution, and Secretion Processes

“…It has been clearly established that neurons in primary cultures can enter a differentiation program (Yavin and Yavin, 1974). Morphologically, these cells are spherical in shape at a nondifferentiated stage and progressively display extended processes that ultimately give rise to a typical network of interconnecting fibers after 24 hr to 4 d in culture.…”

Rapid Assembly of Functional Presynaptic Boutons Triggered by Adhesive Contacts

“…PDL is typically used to coat substrates to promote cell adhesion in vitro (Yavin and Yavin, 1974). In previous studies, it had been reported that isolated axons grown on a PDL-coated substrate form presynaptic-like structures similar in composition to the bead-presynaptic complexes reported here, but these structures were highly mobile and not thought to be triggered by the substrate itself (Krueger et al, 2003).…”

Catecholamine secretion in a rat pheochromocytoma cell line: two pathways for calcium entry.

“…The cells were grown on tissue culture dishes and transferred after vigorous pipetting to remove and dissociate the cells. Experiments were performed on cells which were replated onto polylysine coated (Yavin & Yavin, 1974) tissue culture wells and used within 3 days. Each well, which had a surface area of 80 mm2, contained about 106 cells.…”