Mem basal medium

The animal study was reviewed and approved by the Institutional Animal Care and Use Committee at Northwest A&F University (NWAFU.No20200620c0600025[176]). Healthy female cats (three months old) were used for isolation of adipose mesenchymal stem cells. After anesthesia, the groin adipose tissue was taken aseptically and cut into one mm³ pieces for collagenase digestion. Collagenase (type 1, Gibco) (0.1%) was added to the tissue pieces and incubated at 37 °C for one hour. Then, the supernatants were discarded by centrifuging at 250 g for 5 min. The cell pellets were resuspended with 5 ml MEM basal medium and filtered through a cell strainer. Finally, the cell pellets were collected by centrifugation (250 g, 5 min) again and cultured with ADMSCs medium (88% MEM basal medium (Gibco), 1% penicillin and streptomycin (Gibco), 1% glutamine (Gibco) and 10% fetal bovine serum (Biological Industries)). After 24 h, non-adherent cells were removed, and the cell culture was replaced with the fresh ADMSCs medium. When the ADMSCs grow up to 80% confluence, cells were trypsinized and collected for liquid nitrogen cryopreservation. In this study, ADMSCs within four passages were used for subsequent treatment trials.

HepG2 human liver cells and AML-12 mouse liver cells (Procell, China) were cultured in MEM basal medium (Gibco, China) containing 10% fetal bovine plasma (Gibco, Australia). After reaching a certain order of magnitude in the culture flasks, the cells were inoculated into six-well plates (Corning, China) with 3 × 10⁵ cells per well, and after cell apposition, the cells were treated with KYNA for 2 and 48 h respectively. Cells were then collected to extract RNA or protein.