U 251mg

Human malignant glioma cell lines A-172 (cell no. JCRB0228; lot no. 021999), AM-38 (cell no. IFO50492; lot no. 12082003), T98G (cell no. IFO50303; lot no. 1007), U-251MG (cell no. IFO50288; lot no. 12132002), and YH-13 (cell no. IFO50493; lot no. 1164) were purchased from Health Science Research Resources Bank (Sennan, Osaka, Japan). U-87MG (glioblastoma of unknown origin; cat. no. HTB-14; lot no. 2497162) and U-138MG (cat. no. HTB-16; lot no. 1104428) were purchased from the American Type Culture Collection (Manassas, VA, USA). In a previous study, we confirmed that O⁶-methylguanine-DNA methyltransferase (MGMT, a key factor of alkylating agents) is expressed in T98G, U-138MG and YH-13 cells by real-time RT-PCR and western blot analysis (29 (link)). Consistent with an earlier study (30 (link)), it was also confirmed that T98G (237 Met→Ile) and U-251MG (273 Arg→His) have a point mutation in the p53 gene (data not shown).
Cells were cultured in Dulbecco's modified Eagle's minimum essential medium (DMEM; Nissui Pharmaceutical Co., Ltd.) supplemented with 10% fetal calf serum (FCS; Life Technologies; Thermo Fisher Scientific, Inc.) using plastic culture flasks (Corning, Inc.) in a 37°C-humidified incubator with 5% CO₂. Natural-type IFN-β (Toray Industries, Inc.) and TRAIL (Wako Pure Chemical Industries, Ltd.) were employed for the experiments.

The human malignant glioma cell lines A-172 (cat. no. JCRB0228; lot no. 021999), AM-38 (cat. no. IFO50492; lot no. 12082003), T98G (cat. no. IFO50303; lot no. 1007), U-251MG (cat. no. IFO50288; lot no. 12132002) and YH-13 (cat. no. IFO50493; lot no. 1164) were purchased from the Health Science Research Resources Bank of Japan. U-138MG (cat. no. HTB-16; lot no. 1104428) was purchased from the American Type Culture Collection. A previous study by our group confirmed that O⁶-methylguanine-DNA methyltransferase (MGMT), a key factor of alkylating agents, was expressed in the T98G, U-138MG and YH-13 cell lines via RT-qPCR and western blot analyses (22 (link)). Consistent with an earlier study (23 (link)), it was also confirmed that the T98G (237 Met→Ile) and U-251MG (273 Arg→His) cell lines have a point mutation in the TP53 gene (data not shown).
Cells were cultured in Dulbecco's modified Eagle's medium (Nissui Pharmaceutical) supplemented with 10% fetal calf serum (Thermo Fisher Scientific, Inc.) using plastic culture flasks (Corning, Inc.) in a 37°C incubator in a humidified (>95%) atmosphere containing 5% CO₂.
The antiepileptic drugs perampanel (kindly gifted by Eisai Co., Ltd.), CBZ (Tokyo Chemical Industry), VPA (Tokyo Chemical Industry) and LEV (Tokyo Chemical Industry) and the anti-cancer agent TMZ (Tokyo Chemical Industry) were employed for this study.