Abstract
Dissociated neonatal rat cerebellar cells were grown on medium supplemented with 10% horse serum (HS) and compared with those grown using a serum-free supplemented (SFS) medium, modified from Bottenstein and Sato (1979). containing insulin, transferrin, progesterone, putrescine, and selenium (after an initial 24 hr in 10% horse serum). Cells survived for several weeks using either medium. Cells grown in SFS had higher levels of GABA uptake than cells grown in HS. Cellular morphology and the proportion of neurons to glial cells were similar under the two conditions. Transferrin concentrations of 0.5, 10, and 100 µg/ml were tested. Neither neuronal nor glial cells were sensitive to this 200-fold variation. The SFS medium supports survival and maturation of both neurons and glial cells from rat cerebellum. However, the medium is not completely defined since (1) one day of serum is still required and (2) the heterogeneous cell population is undoubtedly conditioning the medium to some extent.
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References
Bottenstein, J. E., and Sato, G. H. (1979). Growth of a rat neuroblastoma cell line in serum-free supplemented medium.Proc. Natl. Acad. Sci. USA 76514–517.
Bottenstein, J. E., Skaper, S. D., Varon, S. S., and Sato, G. H. (1980). Selective survival of neurons from chick embryo sensory ganglionic dissociates utilizing serum-free supplemented medium.Exp. Cell Res. 125183–190.
Eng. L. F., and Rubenstein, L. J. (1978). Contribution of immunohistochemistry to diagnostic problems of human cerebral tumors.J. Histochem. Cytochem. 26513–522.
Fields, K. L. (1979). Cell-type specific antigens of cells of the central and peripheral nervous system.Curr. Topics Develop. Biol. 13237–257.
Foster, A. C., and Roberts, P. J. (1980). Morphological and biochemical changes in the cerebellum induced by kainic acid in vivo.J. Neurochem. 341191–1200.
Hatten, M. E., and Messer, A. (1978). Postnatal cerebellar cells from staggerer mutant mice express embryonic cell surface characteristic.Nature 276504–506.
Herndon, R. M., and Coyle, J. T. (1977). Selective destruction of neurons by a transmitter agonist.Science 19871–72.
Hökfelt, T., and Ljungdahl, A. (1972). Autoradiographic identification of cerebral and cerebellar cortical neurons accumulating labelled GABA (3H).Exp. Brain Res. 14354–362.
Hösli, E., and Hösli, L. (1978). Autoradiographic localization of the uptake of3H-β-alanine in rat nervous tissue cultures.Experientia 341519–1521.
Iversen, L. L., and Kelly, J. S. (1975). Uptake and metabolism of gammaaminobutyric acid by neuronal and glial cells.Biochem. Pharmacol. 24933–938.
Lowry, O. H., Rosenbrough, N. J., Farr, A. L., and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193265–275.
Mains, R. E., and Patterson, P. H. (1973). Primary cultures of dissociated sympathetic neurons I. Establishment of long-term growth in culture and studies of differentiated properties.J. Cell Biol. 59329–345.
Messer, A. (1977). The maintenance and identification of mouse cerebellar granule cells in monolayer culture.Brain Res. 1301–12.
Messer, A. (1978). Abnormal staggerer cerebellar cell interactions and survival in vitro.Neurosci. Lett. 9185–188.
Messer, A., and Maskin, P. (1980). Short-term effects of kainic acid on rat cerebellar neurons in monolayer cultures.Neurosci. Lett. 19173–178.
Messer, A., Maskin, P., and Mazurkiewicz, J. E. (1980). Effects of using a chemically defined medium for primary rat monolayer cerebellar cultures: morphology, GABA uptake, and kainic acid sensitivity.Brain Res. 184243–247.
Messer, A., and Smith, D. (1977a). In vitro behavior of granule cells from agranular neurological mutants of mice.Brain Res. 13013–23.
Messer, A., and Smith, D. (1977b). Monolayer cultures of mouse cerebellar cells.Tissue Culture Assoc. Manual 3691–693.
Schon, F., and Kelly, J. S. (1975). Selective uptake of3H-β-alanine by glia: association with the glial uptake system for GABA.Brain Res. 86243–257.
Skaper, S. D., Adler, R., and Varon, S. (1979). A procedure for purifying neuronlike cells in cultures from central nervous tissue with a defined medium.Develop. Neurosci. 2233–237.
Tran, V. T., and Snyder, S. H. (1978). Amino acid neurotransmitter candidates in rat cerebellum: selective effects of kainic acid lesions.Brain Res. 167345–353.
Trenkner, E., Hatten, M. E., and Sidman, R. L. (1978). Effect of ether-soluble serum components in vitro on the behavior of immature cerebellar cells in weaver mutant mice.Neuroscience 31093–1100.
Vaccaro, D. E., Messer, A., Dichter, M. and Leeman, S. (1980). Primary cultures of dispersed hypothalamic cells from fetal rats: morphology, electrical activity and peptide contents.J. Neurobiol. 11417–424.
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This work was supported in part by grants from the Scottish Rite Schizophrenia Research Program, N.M.J., USA, and by Biomedical Research Grant S 07-RR5394, from the National Institutes of Health, PHS/DHHS.
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Messer, A., Mazurkiewicz, J.E. & Maskin, P. Growth of dissociated rat cerebellar cells using serum-free supplemented media and varied transferrin concentrations. Cell Mol Neurobiol 1, 99–114 (1981). https://doi.org/10.1007/BF00736042
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DOI: https://doi.org/10.1007/BF00736042