RN46A

Кат. номер

12061302-DNA-5UG

12061302-RNA-5UG

12061302-1VL

Embryonic rat medullary raphe, temperature-sensitive mutant of SV40 large T-antigen, immortalised, serotonergic, neuronal
RN46A , an immortalized serotonergic neuronal cell line, was cloned by serial dilution following infection of dissociated embryonic day 13 rat medullary raphe cells with a retrovirus encoding the temperature-sensitive mutant of SV40 large T-antigen (T-ag), RN46A cells are capable of differentiating at 39 °C the non-permissive temperature. Under differentiation conditions, RN46A cells cease dividing, take on a neuronal morphology, and express enhanced levels of NSE and all three NF proteins. Differentiated RN46A cells express low-affinity nerve growth factor (NGF) receptor (p75NGFR) and are i mMunoreactive using an antibody that recognizes the carboxy-terminal 13 amino acids of all three trk proteins (pan-trk). Both i mMunoreactivities could be potentiated by treatment with brain-derived neurotrophic factor (BDNF), NGF, and adrenocorticotropic hormone, fragment 4-10 (ACTH4-10). Differentiated RN46A cells express low levels of tryptophan hydroxylase (TPH) i mMunoreactivity, which could be enhanced by treatment with ACTH4-10, BDNF, or NGF. Low levels of serotonin i mMunoreactivity are detected in differentiated RN46A cells, and this was potentiated by differentiating RN46A cells with BDNF for 8 d and 40 mM KCl for days 4-8. RN46A cells should prove useful to elucidate intracellular mechanisms that control neurofilament assembly and 5-HT expression in differentiating raphe neurons.
Not specified
DMEM:F12 (1:1), (D8062) + L-Glutamine, (G7513) + 10% FBS / FCS, (F2442) + 0.25mg/ml Geneticin (G418). Alternatively CNS medium can be used (see Kawamoto & Barrett 1986). Differentiation can be induced as follows: cells growing at 33 °C are sub-cultured onto collagen/fibronectin matrix (100 µg/cm2 air-dried collagen I from rat tail followed by 1 µg/cm2 fibronectin). Sub-confluent cells (75%) are shifted to 39 °C. The culture medium is changed to DMEM:F12 (1:1), (D8062) + 1%(w/v) bovine serum albumin (BSA) + 1 μg/ml bovine transferrin + 5 μg/ml bovine insulin + 100 nM putrescine + 20 nM progesterone.
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 19hrs.
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biological source	rat embryo (day 13 medullary raphe)
growth mode	Adherent
karyotype	Not specified
morphology	Fibroblast morphology while proliferating and neuronal on differentiation
receptors	Not specified

RIDADR	NONH for all modes of transport
WGK Germany	3