Human SGPL1 Stable Knockout N/TERT-1 Cell Line

Propagation Media:PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture. Combine 3:1 (v/v) PriGrow IV (TM004): PriGrow III (TM003) + 10% FBS + 5µg/ml Insulin (TM053) + 0.4µg/ml Hydrocortisone (TM066) + 10−10 M cholera enterotoxin + 10ng/ml epidermal growth factor  (Z100139)+ 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. To reduce the differentiated state of cells once they have been propagated in the propagation media, switch to the following media for one passage prior to experimental application: PriGrow X Series Medium for T3421 (TM3421) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. 

This cell line features a stable knockout of Sphingosine 1-phosphate lyase (SGPL1) in nTERT immortalized human keratinocyte using second generation lentivial trasduction. Transduced cells were selected with puromycin and single-cell sorted. Gene knockout was evaluated using Western blot and sequencing data. This cell line is suitable for SGPL1 insufficiency syndrome (SPLIS) related research.