Hek293t

HEK293T and HeLa cells were purchased from the JCRB Cell Bank. Lenti-X 293 T cells were purchased from Takara Bio (Shiga, Japan). NIH3T3, BEAS2B, A549, and HCC827 cells were purchased from American Tissue Culture Collection (ATCC). p53-knockout mouse embryonic fibroblasts (p53^−/− MEFs) were obtained as described elsewhere^{73 (link)}. Mycoplasma contamination was routinely monitored using MycoStrip (InvivoGen, San Diego, CA, USA). HEK293T, HeLa, NIH3T3, and p53^−/− MEFs were cultured in Dulbecco’s Modified Eagle Medium (DMEM; Nacalai Tesque, Kyoto, Japan) supplemented with 10% fetal bovine serum (FBS; Nichirei Biosciences, Tokyo, Japan). Lenti-X 293 T cells were cultured in DMEM (high glucose) supplemented with 10% FBS. BEAS2B cells were cultured in DMEM/F-12K (Nacalai Tesque) supplemented with 10% FBS. A549 and HCC827 cells were cultured in RPMI-1640 medium (Nacalai Tesque) supplemented with 10% FBS. Streptomycin (100 U/ml)/Penicillin-G (100 μg/ml) solution (Nacalai Tesque) was added to all cell culture media.

Human GC cell line (AGS) was purchased from American Type Culture Collection (ATCC, USA). HEK293T and MKN7 cell lines were purchased from the Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan). These cell lines have been tested and authenticated using the STR-PCR method. The highly disseminated peritoneal human GC cell line (As44) and its control cell line (HSC44) were provided by Yanagihara [20 (link), 21 (link)]. The HSC44 cell line was derived from patients with gastric scirrhous cancer. The cell line As44 showing high metastatic potential was derived from HSC44 following 12 cycles of direct orthotropic transplantation to the gastric walls of nude mice and gathering cells from the ascites. These cell lines were cultured in RPMI 1640 (Gibco, CA, USA) supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified atmosphere containing 5% CO₂.

The human chronic myelogenous leukemia cell line K562 and subclones of the human embryonic kidney cell lines HEK293, HEK293T, and the Lenti-X 293 T cell line were obtained from JCRB Cell Bank (Osaka, Japan), RIKEN Bio-Resource Center, and Clontech (Mountain View, CA, USA), respectively. The K562/HER2, K562/HER2/scFv, and K562/HER2/scFv^B cell lines were established through lentiviral transduction of K562 and K562/HER2 cells with the plasmids pCSII/HER2-mCherry, pCSII/scFv-sfGFP, and pCSII/scFv-BFP, respectively.
K562, K562/HER2, K562/HER2/scFv, and K562/HER2/scFv^B cells, as well as HEK293T and Lenti-X 293T cells, were maintained in 10% FBS-RPMI-1640 (Invitrogen, Waltham, MA, USA) and 10% FBS-DMEM (Nacalai Tesque, Kyoto, Japan), respectively. All media were supplemented with penicillin (100 U/mL) and streptomycin (100 μg/mL) (Nacalai Tesque). Cells were maintained in a 37 °C incubator with 5% CO₂ and regularly checked for mycoplasma contamination using a mycoplasma test kit (Lonza, Basel, Switzerland).