RN46A-B14

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.

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.

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).

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