Rpmi1640 medium

Immediately after surgical resection of the tumor, several tissue blocks (approximately 0.5 cm) were removed from the center of the tumor mass to avoid blood vessels. The tissue blocks were placed in RPMI1640 medium (iCell, Shanghai, China), sealed and stored at 4 °C, and transported to the laboratory within 1 h. Tumor tissues were washed three times with D-Hanks solution (Solarbio, Beijing, China) and dissected into 1 mm³ samples in a culture dish. The tissue was mixed with 5 mL type II collagenase (final concentration of 1 mg/mL; Solarbio), followed by digestion for 1.5 h in a 37 °C water bath (Figure 7C). Digestion was terminated by adding 5 mL culture medium and centrifugation at 1000 rpm for 5 min. The cell suspension was filtered twice through a 200-mesh filter. Finally, the RPMI1640 complete medium was added, and the cell suspension was transferred to T25 culture flasks. The complete medium was supplemented with 15% FBS (iCell), 1% penicillin-streptomycin solution (iCell), 1× L-glutamine (iCell), and 20 ng/mL basic fibroblast growth factor (bFGF, iCell). During the culture period, cells were incubated in a humidified incubator at 37 °C with 5% CO₂, and the medium was changed every 2 d. The primary cell line was named PriGIST, and the PriGIST cells were divided into two parts: one was introduced with genes for immortalization, and the other was used as a control.

The human bladder cancer cell line J82 cells were cultured in MEM Alpha Medium (Gibco, USA). The RT4 cells and T24 cells were maintained in McCoy’s 5A Medium (iCell, China), and the 5637 cells were cultured in RPMI1640 Medium (iCell, China). The UMUC3 cells and TCCSUP cells were cultured in MEM (iCell, China), and the SW780 cells were maintained in L15 Medium (iCell, China). The human urothelial cells line SV-HUC-1 cells were cultured in F12K medium (iCell, China). All of the mediums were supplemented with 10% fetal bovine serum (FBS, BI, ISR), 100 μg/mL of penicillin, and 100 U/mL of streptomycin (Invitrogen). All of the examined cells were grown at 37°C with 5% CO₂.
The SCD inhibitor (A939572) utilized in this study was purchased from MedChem Express (HY-50709).