Mrc 5

The NTRK fusion human colorectal cancer cell line KM12SM was kindly gifted by Dr. I. J. Fidler (MD Anderson Cancer Center) to our laboratory in 1999. KM12SM cells, a highly metastatic variant of KM12C cells, have the TPM3‐NTRK1 gene rearrangement. HCC78 cells were obtained from American Type Culture Collection. The PC‐9 cell line with an EGFR exon 19 deletion was obtained from the RIKEN Cell Bank. A925L cells with EML4‐ALK variant 5a (E2:A20) were kindly provided by Dr. H. Uramoto (Kanazawa Medical University). Human lung fibroblasts MRC‐5 and IMR‐90 were obtained from RIKEN Cell Bank. The KM12SM and HCC78 cell lines were cultured in RPMI 1640 with 10% fetal bovine serum (FBS), penicillin (100 U/ml), and streptomycin (10 mg/ml) in a humidified, 5% CO2 incubator at 37°C. The IMR‐90 and MRC‐5 cell lines were maintained in DMEM containing 10% FBS, penicillin (100 U/ml), and streptomycin (10 mg/ml). Entrectinib and capmatinib were obtained from Selleck Chemicals (Houston, TX, USA). Recombinant NRG, HGF, IGF‐1, FGF2, and amphiregulin were purchased from R&D Systems (Minneapolis, MN, USA). Goat anti‐human HGF neutralizing antibody and control goat IgG were obtained from R&D Systems as well.

MRC-5, a human lung fibroblast cell line, was obtained from Riken Cell Bank (Tsukuba, Japan) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics at 37°C in 5% CO₂/95% air. Normal human lung fibroblasts (NHLF) were purchased from Lonza Japan (Tokyo, Japan) and cultured in Fibroblast Media containing human recombinant fibroblast growth factor β, insulin, FBS, and gentamycin/amphotenricin B. Normal mouse lung fibroblasts (NMLF) were prepared from CD1 mice as described previously [12 (link)] and cultured in DMEM supplemented with 10% FBS and antibiotics. Cells were washed twice with phosphate buffered saline (PBS) to remove FBS, cultured in serum-free Opti-MEM (Gibco) for 24 h, and treated with TGF-β1 to induce myofibroblast differentiation.