Hca 7

Human Caco2 and HCA7 intestinal epithelial cell lines were obtained from ATCC or Sigma-Aldrich, Germany, respectively. Caco2 (2 × 10⁴ cells/well) and HCA7 (5 × 10³ cells/well) cells were cultured in Dulbecco’s modified Eagle’s medium (Merck, Darmstadt, Germany) containing 10% Fetal Bovine Serum (FBS) (Merck, Darmstadt, Germany) and 1% penicillin and streptomycin (PAN-BioTech, Aidenbach, Germany) in 96-well plates at 37 °C in a 5% carbon dioxide atmosphere. Sevelamer (Sanofi-Aventis, Frankfurt am Main, Germany), polystyrene sulfonate (MERCK, Darmstadt, Germany), cholestyramine (Ratiopharm, Ulm, Germany), patiromer (Vifor Fresenius Medical Care Renal Pharma, Paris La Défense CEDEX, France), and monosodium urate (Invivogen, San Diego, CA, USA) drugs/crystals were pestled and suspended in D-PBS prior to sonication to generate small crystals using an Ultrasonic Sonifier (Branson) for 5 min. After intestinal epithelial cells were 80% confluent, cells were stimulated with or without the aforementioned crystal types (200 or 1000 µg/mL) for 24 h. Supernatants were collected and stored at −20 °C until further analysis, and cells were harvested for flow cytometry or fluorescence microscopy.

Human colon cancer cell lines HCA‐7, LoVo, SW620, RKO, SW1116 and SW48 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured according to the recommended protocols. In brief, SW48, SW620 and SW1116 cell lines were cultured in flasks containing RPMI‐1640 medium, LoVo cell line was cultured in Ham's F‐12K medium, HAC‐7 cell line was cultured in Dulbecco's modified Eagle's medium, and RKO cell line was cultured in Eagle's minimal essential medium. All culture media were supplemented with 10% FBS (Gibco, Grand Island, NY, USA). These CRC cell lines were selected as they express different levels of six validated genes (CEA, EpCAM,CK19,MUC1,EGFR and C‐Met).

The human colorectal carcinoma-derived cell lines HCT116, HCA7 and SW480 were obtained from the American Type Culture Collection (ATCC, Maryland, USA) and maintained as previously described.36 (link) HCT116 cells were cultured in McCoy's 5A Medium (Invitrogen, UK), whereas SW480 and HCA7 cells were grown in Dulbecco's Modified Eagle's Medium (DMEM; Source Bioscience, UK), both supplemented with 10% fetal bovine serum (FBS), 2 mM glutamine (Sigma, UK), 100 U/mL penicillin and 100 μg/mL streptomycin (Invitrogen). The non-tumorigenic adenoma-derived clonogenic S/RG/C2 cell line (referred to as RG/C237 (link)
38 (link)) was maintained in DMEM supplemented with 20% FBS, 2 mM L-glutamine (Sigma), 100 U/mL penicillin and 100 μg/mL streptomycin (Invitrogen) as well as 0.2 U/mL insulin (Actrapid Novo Nordisk, Denmark) and 1 μg/mL hydrocortisone (Sigma).