A procedure for culturing rat neocortex explants in a serum-free nutrient medium.
A procedure is described for long-term culturing of rat neocortex explants in a serum-free growth medium. Slices spanning the entire cortical depth from pial to ventricular side are prepared from 6-day-old rat pups. After preincubation in Hanks' balanced salt solution with extra glucose, the explants are placed on polyamide gauze carriers in plastic culture dishes containing serum-free medium. The dishes are continuously rocked during the culture period. After 3 weeks in vitro the explants consist of a three-dimensional network of neural tissue with a mean thickness above the gauze of ca. 100 micron which corresponds with about 8 cell layers. Central necrosis is either fully absent (in one-third of the explants) or restricted to a minimal strip or patch located close to the gauze. From pial to ventricular side, 5 layers can be distinguished which, with respect to cell size and cell density, reflect a histiotypic architecture. The dense neuropil shows abundant axo-dendritic synapses (both on shafts and spines), myelinated fibers, and spontaneous bioelectric activity.
Romijn
H J
HJ
Netherlands Institute for Brain Research, Amsterdam.
de Jong
B M
BM
Ruijter
J M
JM
D000818:Animals; D002540:Cerebral Cortex / Q000166:cytology* / Q000648:ultrastructure; D003470:Culture Media; D007202:Indicators and Reagents; D008854:Microscopy, Electron; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats; D011919:Rats, Inbred Strains
D003470:Culture Media; D007202:Indicators and Reagents