Immortalized Mouse Enteric Neuronal Cells were isolated from a transgenic mouse with a mutation of the SV-40 tsA58 gene. These novel cells are very stable- biologically and phenotypically- for multiple generations, in vitro. Thus, they are perfectly suited for research focusing on the enteric nervous system; from determining the impact of disease on the enteric nervous system to simply tracking its development. Immortalization Method: Isolation from transgenic mouse with SV-40 large T antigen mutationBioSafety Level: IISpecies: MouseSource Organ: Intestine of E13 Mouse Fetus Growth Properties: AdherentMorphology: Epithelial-likePopulation Doubling: 25-35 hoursSeeding Density: 20,000 - 50,000 cells/cm²; Recommended split ratio is 1:2 to 1:6Markers: Peripherin, HuD, PGP9.5, tau, synaptophysin, characteristic enteric neuron receptors, cRet, nestinDonor Age: N/ADonor Gender: N/ADonor Ethnicity: N/APropagation: The base medium for this cell line is Prigrow IV available from abm (TM004). To make the complete growth medium, add the following components to the base medium: 52 ng/ml selenium (Sigma), 16.11 µg/ml putrescine (Sigma), 5 µg/ml insulin (Sigma), 1 µg/ml transferrin (Sigma), 0.1 mg/ml fetuin (Sigma, Cat No. F3385), 1 µg/ml BSA (Bio Basic), 6.3 ng/ml progesterone (Sigma), and L-glutamine (G275) to a final concentration of 1%.Filter the media with 0.45 µM filter before proceeding to add fetal bovine serum (TM999)* to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, 10 ng/ml mouse IFN-gamma (Z200085), and 100 ng/ml glial cell line-derived neurotrophic factor (GDNF) (Z101057) to make complete growth media. Carbon dioxide (CO2): 5%, Temperature: 33.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.Quality Control: Neuronal markers were assessed and confirmed via western blot and immunocytochemical analysis. Shipping Condition: Dry IceStorage Condition: liquid nitrogen or -180°C