Atcc htb 77

Human NIH:OVCAR-3 ovarian cancer cells (ATCC^® #HTB-161™) were commercially obtained in passage 76 (P76) from the Institute for Applied Cell Culture (IAZ), Munich, Germany. The SK-OV-3 ovarian cancer cells (ATCC^® #HTB-77™) were commercially obtained in P25 from the ATCC, Manassas, VA, USA. These two cell lines were originally established from the malignant ascites of a patient with progressive adenocarcinoma of the ovary, respectively. Both cell types were cultivated at about 1,750 cells/cm² in RPMI 1640 supplemented with 10% (v/v) fetal calf serum, 100 U/ml L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin. Subculture was performed by trypsin/EDTA (Biochrom GmbH, Berlin, Germany) treatment for 5 min at 37°C.

The human OV-90 cell line with mutated TP53 gene (human malignant papillary serous carcinoma, American Type Culture Collection (ATCC) CRL-11732™) and SKOV-3 cell line with loss of TP53 function (TP53 null) (human ovarian adenocarcinoma, ATCC HTB-77) were purchased from ATCC (Rockville, MD, USA). BRCA^MUT PEO-1 cells (human ovarian cancer; estrogen rec, 10032308) were obtained from the European Collection of Authenticated Cell Cultures. The newly acquired cells were expanded, and aliquots of less than 10 passages were stored in liquid nitrogen. All cell lines were kept at a low passage, returning to original frozen stocks every 6 months. During the course of the study, cells were thawed and passaged within 2 months of each experiment. Cells were cultured in DMEM and RPMI with 10% FBS and regularly checked for mycoplasma contamination. Cells were cultured in an atmosphere of 5% CO₂ and 95% air at 37 °C.

The HT29 (ATCC^® HTB-38) and CT26 colon (ATCC^® CRL-2638^™), A549 lung (ATCC^® CCL-185^™), SKOV-3 ovarian (ATCC^® HTB-77^™), B16 melanoma (ATCC^® CRL-6323^™), ZR-75-1 breast (ATCC^® CRL-1500^™) and U937 leukemic (ATCC^® CRL-1593.2^™) cell lines used in this study were obtained from the American Type Culture Collection (ATCC^®). MC38 cells were acquired from Kerafast (Boston, MA, USA), where they are authenticated, characterised and certified as mycoplasma free. Secondarily, stored cultures are routinely tested for mycoplasma contamination by PCR.

Human ovarian cancer SKOV-3 (ATCC® HTB-77™) and OVCAR-3 (ATCC® HTB-161™) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) by the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). These cell lines were tested and authenticated by the BCRC. We purchased them from BCRC and passaged them for less than 6 months after thawing and maintained them for further study in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) (for SKOV-3 cells) or with 20% FBS/0.01 mg/ml bovine insulin (for OVCAR-3 cells). All cell cultures were maintained in a 5% CO₂/95% air incubator at 37 °C. prior to treatment, cells were placed in 0.25% hormone-stripped FBS-containing medium for 2 days.

HOFs were obtained from ScienCell (Cat# 7330-SC) and cultured in Fibroblast Medium (Cat# 2301-SC; ScienCell). CAFs were obtained from Vitro Biopharma (Cat# CAF02) and cultured in Low-Serum, VitroPlus III, Complete Medium (Cat# PC00B1). Omental-derived adult primary MCs were obtained from Zen-Bio (USA) and cultured in Mesothelial Cell Growth Medium (Cat# MSO-1; Zen-Bio). The SOC cell line SKOV3 (ATCC® HTB-77™) provided by the American Type Culture Collection (LGC Standards, Teddington, UK), was maintained in McCoy’s 5A medium (Cat# 26600-023; GIBCO Thermo Fisher). OVCA433 cell line was provided by Prof. G. Scambia (Catholic University School of Medicine) and maintained in Dulbecco’s modified Eagle medium (Cat# 21885-025; GIBCO Thermo Fisher). Kuramochi cell line was provided by the National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN, Osaka, Japan), and the OVCAR3 cells (ATCC® HTB-161 were obtained from the American Type Culture Collection (LGC Standards, Teddington, UK), both cultured in RPMI-1640 medium (Cat# 618700-010; GIBCO Thermo Fisher). All media were supplemented with 10% or 20% FBS, containing penicillin (10,000 U/ml)-streptomycin (10 mg/ml). Cells were incubated at 37 °C in a humidified atmosphere containing 5% CO₂ and were tested for the absence of viral/mycoplasma contamination.