Isolation and characterisation of fetal bovine brain cells in primary culture

Primary cultures and cryopreservation procedure of bovine brain cells were established as in vitro experimental systems to study the responses of bovi ne brain cells to neuropathogenic agents. Brain cells were dissociated by p apain from the cerebellum of a bovine fetus at 90 to 120 days old, and were cultured in different media. In a medium containing 1 per cent fetal bovin e serum (FBS), neuronal cells were maintained and they formed clusters on g lial and fibroblastic cell sheets. In a medium containing 10 per cent FBS, the proportion of neurones decreased, and fibroblastic and microglial cells dominated. In a serum-free medium containing epidermal growth factor; the highest neuronal proportion was obtained. Optimal cryopreservation conditio n for the brain tissues was investigated by changing the concentrations of DMSO and FBS. Brain cells could he cultured from cryopreserved tissue with only slightly reduced growth profiles and varying cell proportions in compa rison to those prepared from fresh tissue. (C) 2000 Harcourt Publishers Ltd .