biological source
rat embryo (day 13 medullary raphe)
packaging
tube of 5 μg 12061303-DNA-5UG, pkg of vial of cells 12061303-1VL
growth mode
Adherent
karyotype
Not specified
morphology
Fibroblast morphology while proliferating and neuronal on differentiation
receptors
Not specified
Biochem/physiol Actions
Embryonic rat medullary raphe, temperature-sensitive mutant of SV40 large T-antigen, immortalised, serotonergic, neuronal
Not specified
The clonal cell line, RN46A-B14, was isolated following transfection of the gene encoding rat brain-derived neurotrophic factor (BDNF) into RN46A cells (see ECACC Catalogue number 12061302). RN46A-B14 cells synthesize and secrete biologically active BDNF in vitro and synthesize serotonin (5-HT) following partial membrane depolarization. Two weeks following RN46A-B14 cell transplantation into the adult rat cortex and hippocampus, there is a threefold increase in survival of RN46A-B14 cells compared to RN46A cells. The grafted RN46A-B14 cells i mMunohistochemically stain for BDNF and 5-HT, while RN46A cells transfected with vector only are negative for both BDNF and 5HT. In addition, RN46A-B14 cells attain more morphologically complex phenotypes, indicating enhanced neuronal differentiation. Autocrine secretion of BDNF by RN46A-B14 cells thus potentiates survival and can be used to deliver both BDNF and 5-HT in vivo.
Preparation Note
DMEM:F12 (1:1) (D8062) + L-Glutamine (G7513) + 10% FBS / FCS (F2442) + 0.25mg/ml Geneticin (G418) + 0.1mg/ml hygromycin. Alternatively CNS medium can be used (see Kawamoto & Barrett 1986).
Split subconfluent cultures (70-80%) 1:2 to 1:5 using 0.25% trypsin/EDTA; 5% CO2; 33 °C. Suggested seeding density 2-4 x 10,000 cells/cm2. Doubling time 25hrs.
Other Notes
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This cell line is part of the European Collection of Authenticated Cell Cultures (ECACC), an international repository managed by the United Kingdom Health Security Agency (UKHSA). No licensing agreement is required when either this cell line or the DNA extracted from it are used for internal research purposes only. Any other use of these products is prohibited without the express written permission of UKHSA. Inquiries regarding authorized use of this cell line or its genomic DNA may be directed to culturecollections@ukhsa.gov.uk.