Snu 349

A total of 16 primary RCC patient tumour samples (Table I) and 5 primary RCC cell were kindly provided by Dr Helén Nilsson, Department of Translational Medicine, Lund University for validation of selected biomarkers. Patient tumour samples were retrieved from the material collection established after Lund University ethical committee approval (approval no. LU680-08), where informed written consent was obtained before archival. Human renal tissue dissociation and culturing was performed as described in Appendix S1 and previously described in Hansson et al (23 (link)). Tumour tissues obtained consisted of ccRCC, p1RCC, p2RCC and chRCC tissues and cells that were predetermined by a trained pathologist. ccRCC cell lines SNU-349 (Korean Cell Line Bank) and 786-O (ATCC) were cultured in RPMI-1640 (Corning, Manassas, USA) and DMEM (Corning, Manassas, USA) respectively, both medium supplemented with 10% foetal bovine serum (Gibco, MA, USA). All cell lines were incubated in a humidified chamber in 5% CO₂ at 37˚C, were STR authenticated, had a passage number of not more than 20 and were mycoplasma free. Healthy blood for controls were obtained from the Blood donation centre in Lund with informed written consent from patients and ethical permission obtained by the regional health care provider (Region Skåne, Lund, Sweden; approval no. 2018:19).

Human kidney proximal tubule cells (HK-2) and RCC cells (Caki-1, Caki-2, SNU-333, SNU-349, and SNU-1272) were purchased from the Korean Cell Line Bank, and cultured according to the supplier’s protocol. HK-2, SNU-333, SNU-349, and SNU-1272 cells were cultured in RPMI-1640 medium, and Caki-1 and Caki-2 cells were cultured in DMEM/high glucose. All media were from Welgene and contained 10% fetal bovine serum and 1% penicillin-streptomycin. The cells were grown in monolayers at 37 °C in a 5% CO₂ incubator unless otherwise described.