Nih3t3 cells

NIH3T3 cells were purchased from Health Science Research Resources Bank (Osaka, Japan). HEK293T cells were a kind gift from Dr. Kunio Shiota (The University of Tokyo, Tokyo, Japan). HEK293 cells were kindly provided by Dr. Koichi Suzuki (Teikyo University, Tokyo, Japan). NIH3T3 cells, HEK293T cells, and HEK293 cells were cultured in DMEM containing 1 mg/ml NaHCO₃, 50 IU/ml penicillin, 50 μg/ml streptomycin, 0.5 μg/ml amphotericin B, and 100 μg/ml kanamycin supplemented with 10% FBS (NIH3T3 cells and HEK293 cells) or 10% CS (HEK293T cells). FRTL-5 rat thyroid follicular cells (28 (link)) were kindly provided by the late Dr. Leonard Kohn (Ohio University and Edison Biotechnology Institute, Athens, OH, USA). FRTL-5 cells were cultured as previously described (12 (link)).

We used NIH 3T3 cells (JCRB0615; Health Science Research Resources Bank, Japan Health Science Foundation, Japan) and HeLa cells (JCRB9004; Health Science Research Resources Bank, Japan Health Science Foundation, Japan) as adhesive cells. The cells were cultured in Dulbecco’s minimal essential medium (Invitrogen Corp., USA) supplemented with 10% fetal bovine serum, 100 U mL⁻¹ penicillin, and 100 µg mL⁻¹ streptomycin. Each cell suspension was obtained by treating the confluent monolayer formed on the tissue culture dish with 0.25% trypsin (Invitrogen). The cells were cultured under a humidified atmosphere of 5% CO₂ and 95% air at 37 °C.