Dmem f12

The Human Embryonic Kidney cell line (HEK293T) was grown in DMEM high glucose (Wisent, St-Bruno, Canada) containing 10% FBS, 2 mM L-glutamine and penicillin/streptomycin. The ER+ human breast adenocarcinoma cells, MCF7, were cultured in DMEM/F12 (Wisent Bioproducts, St-Bruno, Quebec, Canada) supplemented with 5% FBS, 2 mM L-glutamine, penicillin/streptomycin, 15 mM HEPES, 0,2% sodium bicarbonate and 10E-9 M estradiol. The human cervical carcinoma cells, HeLa, were grown in DMEM medium (Wisent) with 5% FBS, 2 mM L-glutamine and penicillin/streptomycin. Cells were grown at 37°C in a 5% CO2 incubator.

Mouse mesenchymal stem cells and dendritic cells were used in the present study. MSCs derived from the bone marrow of C57BL/6 mice were purchased from Cyagen Biosciences Inc. (Guangzhou, China). The supplier identified MSCs based on the cell surface phenotype and pluripotency. MSCs were cultured in DMEM/F12 containing 10% fetal bovine serum (Wisent, Nanjing, China) and grown in a humidified 5% CO₂ sterile incubator at 37 °C.

The human EOC cell lines [TOV81D, TOV112D, TOV1946, OV866(2), OV1946, TOV2295(R), OV4485] were derived in our laboratory from patients’ tumors (TOV) or ascites (OV) and characterized in detail [34 (link)–38 (link)]. All EOC cell lines were maintained in a low oxygen condition of 7% O₂ and 5% CO₂ and grown in OSE medium (Wisent, Montreal, QC). The human retinal epithelial cell line ARPE was purchased from American Type Culture Collection (ARPE-19, #CRL2302) and maintained in DMEM-F12 (Wisent). All media were supplemented with 10% FBS (Wisent), 0.5 μg/mL amphotericin B (Wisent), and 50 μg/mL gentamicin (Life Technologies Inc). Antibodies, chemicals, oligonucleotides, plasmids, and commercial assays used in this study are indicated in Supplementary Table S1.

The human OC cell lines CaOV3 and OVCAR3 were obtained from the American Type Culture Collection (Manassas, VA) and maintained in a humidified 5 % CO₂ incubator at 37 °C. Cells were passaged twice weekly. OVCAR3 cells were maintained in RPMI-1640 (Wisent, St-Bruno, QC, Canada) supplemented with 20 % FBS, insulin (10 mg/L), glutamine (2 mM) and antibiotics. CaOV3 cells were cultured in DMEM/F12 (Wisent) supplemented with 10 % FBS, 2 mM glutamine and antibiotics. Acellular ascites fractions were obtained at the time of initial cytoreductive surgery from women with advanced serous ovarian carcinomas. Samples were supplied by the Banque de tissus et de données cliniques et biologiques sur les cancers gynécologiques et du sein de Sherbrooke as part of the Banque de tissus et de données du Réseau de Recherche en Cancer des Fonds de Recherche du Québec en Santé (FRQS) affiliated to the Canadian Tumor Repository Network (CTRNet). HRP-conjugated anti-mouse and rabbit antibodies and anti-FAK antibody were purchased from Cell Signaling Technology (Danvers, MA). Anti-phospho FAK and anti-phospho Pyl2 were from Thermo Fisher (Waltham, MA). Anti-Pyk2 and anti-Tubulin were purchased from Sigma-Aldrich (Oakville, ON). CCL18 and CCL18 neutralizing antibody were from RnD Systems (Minneapolis, MN). Plasmid pCMV6-ENTRY-PTK2B was obtained from Origene (Rockville, MD).

POPG (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol), POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and NBD-PC were from Avanti Polar Lipids (Alabaster, AL, USA); Bio-Beads SM2 Adsorbent, Bio-Spin chromatography columns and Bio-Gel P-6 were from Bio-Rad (Hercules, CA, USA); Micro-Spin columns with screw caps were from Thermo Scientific (Rockford, IL, USA); 100 × protease inhibitor cocktail was from EMD Millipore (Billerica, MA, USA); DDM was from Anatrace (Maumee, OH, USA); anti-FLAG M2 agarose and 3 × FLAG peptide were from Sigma (St Louis, MO, USA); SNAP-capture resin was from New England Biolabs, Ipswich, MA, USA); PfuUltra II DNA polymerase was from Agilent (Santa Clara, CA, USA); QiaFilter Plasmid Maxi kit was from Qiagen (Valencia, CA, USA); DMEM, 100 × penicillin/streptomycin were from Invitrogen (Grand Island, NY, USA); heat-inactivated fetal bovine serum (FBS) was from Atlanta Biologicals (Lawrenceville, GA, USA); DMEM/F12 was from Wisent (Saint Bruno, QC, Canada); FBS and DPBS from Life Technologies (Waltham, MA, USA); doxycycline from Bio Basic (Markham, ON, Canada); blasticidin and puromycin from Bioshop Canada (Burlington, ON, Canada); Jetprime transfection reagent from Polyplus Transfections (Illkirch, France); and Miniprep kit from Qiagen.