National Institute of Animal Health (NIAH)

Topics in Animal Health Research 2000

23. Establishment of Primary Cell Culture from Fresh and Cryopreserved Fetal Bovine Brain Tissue


  Procedures for primary cultures and cryopreservation 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 bovine fetuses at 90-120 days of age, and were cultured in different media. In medium containing 1% fetal bovine serum (FBS), neuronal cells were maintained and they formed clusters on glial and fibroblastic cell sheets. In medium containing 10% FBS, the proportion of neurons decreased, and fibroblastic and microglial cells dominated. In serum-free medium containing epidermal growth factor, the highest neuronal proportion was obtained. Optimal cryoperservation conditions for the brain tissues were 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 Biology and Immunology, National Institute of Agrobiological Sciences (NIAS) Tel:+81-298-38-7801)


  • A. Hashimoto, T. Onodera, H. Ikeda and H. Kitani: Isolation and characterisation of fetal bovine brain cells in primary culture. Res. Vet. Sci. 69: 39-46 (2000)
  • I. Yamane, H. Kitani, T. Kokuho, T. Shibahara , M. Haritani, T. Hamaoka, S. Shimizu, M. Koiwai, K. Shimura and Y. Yokomizo: The inhibitory effect of interferon gamma and tumor necrosis factor alpha on intracellular multiplication of Neospora caninum in primary bovine brain cells. J. Vet. Med. Sci. 62: 347-351 (2000)
  • H. Kitani, M. Yamakawa and H. Ikeda: Preferential infection of neuronal and astroglia cells by Akabane virus in primary cultures of fetal bovine brain. Vet. Microbiol. 73: 269-279 (2000)