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 bovine brain cells to neuropathogenic agents. Brain cells were dissociated by papain 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 bovine serum (FBS), neuronal cells were maintained and they formed clusters on glial 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 condition for the brain tissues was investigated by changing the concentrations of DMSO and FBS. Brain cells could be cultured from cryopreserved tissue with only slightly reduced growth profiles and varying cell proportions in comparison to those prepared from fresh tissue.
Department of Molecular Immunology, Faculty of Agriculture, University of Tokyo, Japan.
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Hashimoto
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Onodera
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Ikeda
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Kitani
D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000737:chemistry; D015214:Antigens, Differentiation, Myelomonocytic / Q000737:chemistry; D001253:Astrocytes / Q000166:cytology; D001921:Brain / Q000166:cytology*; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000662:veterinary; D002525:Cerebellar Cortex / Q000166:cytology; D015925:Cryopreservation / Q000662:veterinary; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000737:chemistry; D005312:Fetal Blood; D005347:Fibroblasts / Q000166:cytology; D005904:Glial Fibrillary Acidic Protein / Q000737:chemistry; D007150:Immunohistochemistry; D008858:Microscopy, Phase-Contrast / Q000662:veterinary; D008869:Microtubule-Associated Proteins / Q000737:chemistry; D009474:Neurons / Q000166:cytology; D010206:Papain / Q000737:chemistry; D000097382:CD68 Molecule
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D003470:Culture Media; D004815:Epidermal Growth Factor; D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D010206:Papain; C067980:CD68 antigen, human; D000097382:CD68 Molecule