Pc3 crl 1435

PC‐3 (CRL‐1435) and DU145 (HTB‐81) human prostate cancer cell lines were purchased from ATCC (Manassas, VA) and maintained in the recommended culture media. The identity of the cell line was checked by Cell ID System and Promega GenePrint 10 System through short tandem repeat analysis (Mission Biotech, Taipei, Taiwan), and passage numbers used for the experiments are <25. A comprehensive methods section is available in Supplementary methods.

Human prostate cancer cell lines (PC3, CRL-1435; DU145, HTB-81; LNCaP, CRL-1740) were obtained from the American Type Culture Collection (Rockville, MD). The metastatic subclone of LNCaP, C4-2B, was a subline derived from bone metastasis of LNCaP bearing-mouse. Luciferase-expressing prostate cancer cells were established by lentiviral transduction. Murine osteoblast cells were established as previously reported [2 (link)]. All prostate cancer cell lines were routinely grown in RPMI 1640 (11875–093, Life Technologies), and murine osteoblast cells were grown in α-MEM (12571–063, Life Technologies). Cultures were supplemented with 10% (v/v) fetal bovine serum (900–108, GEMINI Bio-Products, Sacramento, CA), 1% (v/v) penicillin-streptomycin (15140–122, Life Technologies) and maintained at 37°C, 5% CO₂, and 100% humidity. These cells were certified by DDC Medical.

Prostate cancer cell lines LNCaP (CRL-1740) and PC-3 (CRL-1435) were acquired from the American Type Culture Collection (ATCC, Inc., Manassas, VA 20110-2209 USA) [70 ,71 ]. All cells were cultured with RPMI 1640 media (pH = 7.4) (Sigma–Aldrich, Inc., St. Louis, MI, USA, R8758) [72 ] with 10% of fetal bovine serum (FBS) and incubated under standard conditions (5% CO₂, 37 °C).

Human prostate cancer cell lines PC-3 (CRL-1435) and LNCaP (CRL-1740) were obtained from the American Type Culture Collection (ATCC, United States). Cells were cultured in DMEM:F12 medium (1 : 1) supplemented with 10% fetal calf serum (HyClone, United States), 0.01 g/L gentamicin, 0.3% amphotericin B, and 584 g/L L-glutamine (PanEco, Russia). Passaging was performed every 3 days in a ratio of 1 : 5 using a 0.05% trypsin-EDTA solution (PanEko, Russia).

The HSC-2 (JCRB0622), HSC-3 (JCRB0623), and HSC-4 (JCRB0624) human head and neck squamous cell carcinoma (HNSCC) cell lines were obtained from the Japanese Collection of Research Bioresources (JCRB) cell bank (Osaka, Japan). Human gingival fibroblasts (HGF, CRL-1740) and the PC3 (CRL-1435) and LNCaP (CRL-1740) human prostate cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells used in this study were maintained in Dulbecco’s modified Eagle’s medium (Sigma, St. Louis, MO, USA) supplemented with 10% fetal bovine serum (FBS; Invitrogen, Carlsbad, CA, USA; complete DMEM) at 37 °C in a humidified atmosphere containing 5% CO₂. The establishment of RANKL-expressing HSC-2 HNSCC cells was described previously18 (link). Among them, the RANKL-expressing R1 and R2 cells and the control C1 cells were used in this study.

Human PCa DU145 (HTB-81) and PC3 (CRL-1435) cell lines was obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and was received on 2016/2018. This cell line has been tested and authenticated by DNA fingerprinting by the ATCC. After reception, cells were amplified in order to make a large reserve of cryopreserved cells. Every 3 months, a new cryopreserved bulb was thawed and used for this study. DU145 and PC3 cell lines were grown in RPMI medium (BE12-702F, Lonza, Levallois-Perret, France), supplemented with 5% FBS (CVFSVF0001, Eurobio, Les Ulis, France) and 1% (v/v) penicillin–streptomycin in a humidified incubator at 37 °C with 5% CO₂. All experiments were performed with mycoplasma-free cells. DU145 and PC3 PCa cell lines have been chosen for their high migration capacity.
LA (L1876), EPA (17266), palmitic acid (PA) (P5177), and TGFβ1 (H8541) were from Sigma-Aldrich. GSK7975A (GLXC03243) and Synta66 (GLXC03244) were from GLXX Lab IMC. 1-Ohexadecyl-2-O-methyl-sn-glycero-3-lactose (Ohmline) was synthetized as previously described [33 (link)].