Mem e

The mouse monocyte/macrophage-like cell line RAW 264.7 and human foreskin fibroblasts (BJ cell line) were obtained from the American Type Culture Collection. The RAW 264.7 and BJ cells were cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) and Minimum Essential Medium (Eagle) [MEM(E)] (HiMedia Laboratories, LLC) media, respectively. The culture media were supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and 100 units of penicillin/100 µg/ml streptomycin (HiMedia Laboratories, LLC), and the cells were grown in a humidified incubator in an atmosphere with 5% CO₂ at 37˚C.

Fibroblasts and macrophages were used as a model for inflammatory cells present in hemorrhoids and fistulas. The human foreskin fibroblast cell line BJ and the mouse monocyte/macrophage-like cell line RAW 264.7 were purchased from the American Type Culture Collection. BJ and RAW 264.7 cells were cultured in MEM(E) (HiMedia Laboratories LLC) and RPMI (Gibco; Thermo Fisher Scientific, Inc.) media respectively, supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and 100 units of penicillin and 100 µg/ml streptomycin at 37°C in a humidified atmosphere with 5% CO₂. All treatments were performed in complete medium.