Fetal bovine serum (fbs)

All 60 breast cancer tumor and adjacent non-tumor samples from patients were recruited from the Huaihe Hospital in Kaifeng, China. This study was approved by the ethics committee of Medical School, Henan University. Human breast cancer cell line MCF7 and Mammary epithelial cell lines (Hs578Bst and MCF 10A) were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), and cultivated in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Zeta-life, USA) in a 37°C incubator with 5% CO₂.

The method of culturing bovine hepatocytes has been described in previous studies [11 (link),12 (link)]. Briefly, the donors were healthy female calves purchased from a commercial dairy farm (Hefei, China). Calves were stunned and left decubitus by pentobarbital sodium (0.1 mL/kg body weight). An incision of about 20 cm was made in the right abdomen to cut through the skin, muscle layer and peritoneum. A retractor is placed inside the incision. The caudate lobe of the liver is obtained using a scalpel. Following perfusion cleansing of the obtained tissue, a two-step collagenase IV infusion was used to break down the caudate lobe [13 (link)]. After filtering, the suspension was centrifuged at 500×g for 5 min at 4°C. The cells were then revived in Dulbecco’s modified eagle medium (DMEM) with low glucose (Biosharp, Beijing, China) supplemented with 10% fetal bovine serum (Zeta Life, Menlo Park, CA, USA), 1 nM dexamethasone, 10 μg/mL vitamin C, and 1% antibiotic solution (Invitrogen, Carlsbad, CA, USA; streptomycin, 5×penicillin, amphotericin). The media was changed to DMEM with 10% fetal bovine serum and 1% antibiotic solution following 4 h of culture.
In 6-well plates, 1×10⁶ cells were seeded in each well for 44 h. For the next 12 h, DMEM media containing 0, 1.25, 2.50, 3.75, and 5.00 mM sodium propionate were employed.

Canine pancreatic islets were isolated by collagenase V (Sigma-Aldrich, Burlington, MA, United States) digestion and purified by centrifugation on a Ficoll density gradient (Borot et al., 2013 (link)). Purified islets were incubated (37°C, 5% CO₂) in DMEM medium (Gibco, Waltham, MA, United States) supplemented with 10% fetal bovine serum (Zeta Life, Menlo Park, CA, United States), 100 U/mL penicillin (Sigma, Ronkonkoma, NY, United States), 0.1 mg/mL streptomycin (Sigma, Ronkonkoma, NY, United States), and 0.5 μg/mL Mycoplasma Removal Agent (MP Biomedicals, Irvine, CA, United States).

Grass carp ovary cell line (CO) were kindly provided by Zhejiang institute of freshwater fisheries (Huzhou, Zhejiang, China) and cultured at 28 ± 0.5 °C in humidified atmosphere with 5% CO₂, and maintained in DMEM (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (ZETA LIFE, St. Louis, MO, USA); Grass carp kidney cells (CIK) cells (kindly provided by Prof. Ling-bing Zeng in Yangtze River Fisheries Research Institute, Wuhan, Hubei, China) were cultured at 25 ± 0.5 °C in humidified atmosphere with 5% CO₂, and maintained in Medium 199 (Hyclone, St. Louis, MO, USA) supplemented with 10% fetal bovine serum; Grass carp macrophage was separated from grass carp head kidney by using a fish tissue mononuclear cell separation kit (Solarbio, Beijing, China).

ORECs are provided by iCell Bioscience Inc. (Shanghai, China) and were identified by immunohistochemistry. Cells were cultured in DMEM/F12 containing 10% fetal bovine serum (ZETA LIFE, Menlo Park, CA, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco, Shanghai, China) at 37 °C in 5% carbon dioxide.

In this study, human myeloma cells (RPMI-8226 and U266) and human renal epithelial cells (293T) were obtained from the Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy. All the cells were cultured in RPMI-1640 medium (KeyGEN BioTECH) containing 10% fetal bovine serum (ZETA LIFE) at 37 °C with 5% CO₂ in the incubator.