Rpmi 1640 medium

All cell models were kept under humidified conditions containing 5% CO2 at 37 °C (normal cell culture conditions) and were regularly checked for mycoplasma contamination. Cell authentication was performed by short tandem repeat (STR) analysis. All primary glioma cell lines originating from the Department of Neurosurgery, Neuromed Campus, Kepler University Hospital, Linz (BTL53, BTL1333, BTL1304, BTL2231, BTL2176) and from the Medical University of Vienna (VBT4, VBT92, VBT125, VBT150, VBT172) were cultured in RPMI-1640 medium (Sigma-Aldrich, Missouri, USA) supplemented with 10% fetal calf serum (FCS, Gibco, Thermo Fisher Scientific, MA, USA).
NMC-G1, and AM38 cells were purchased from the Japanese Collection of Research Bioresources Cell Bank (Japan) and were cultured according to the distributor’s recommendations. DBTRG-05MG was purchased from the “Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH” (Braunschweig, Germany) and cultured in RPMI-1640 medium supplemented with 10% FCS. Neither antibiotics nor any other anti-microbial substances were used during this study. All experiments with both primary and stable cell models were performed between passages 5 and 15.

DK-MG (ACC 277) and Jurkat (ACC 282) were purchased from DSMZ and cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal calf serum (FCS), 100 U/mL Penicillin/Streptomycin, and 2 mM l-glutamine, herein referred to as complete RPMI medium (all from Life Technologies). F98 cells (CRL-2397), F98^EGFRvIII (CRL-2949), and F98^EGFR (CRL-2948) were purchased from ATCC, A431 cells were obtained from Dr. G. Moldenhauer (DKFZ Heidelberg), and cultured in DMEM supplemented with 10% FCS, 100 U/mL Penicillin/Streptomycin, and 2 mM l-glutamine. EGFRvIII- or EGFR-transfected F98 cell cultures were supplemented with 0.2 mg/mL Geneticin (G-418) to maintain stable antigen expression.
Flp-In CHO cells were purchased from Life Technologies and adapted to suspension growth in HyClone CDM4 CHO medium (GE Healthcare) supplemented with 2 mM l-glutamine, HT supplement, and 100 U/mL Penicillin/Streptomycin as well as 100 µg/mL Zeocin (all from Life Technologies) prior to transfection.
All cells were cultured under standard conditions at 37°C, 5% CO₂, and subcultured according to standard protocols.
PBMCs were isolated from healthy volunteers’ buffy coat (German Red Cross, Mannheim, Germany) and T-cells enriched as previously described (26 (link)).

Dami, HEL and Meg-01 cells were purchased from ATCC (ATCC number: CRL-9792, TIB-180 and CRL-2021, respectively) and grown in RPMI 1640 medium (Life Technologies, Grand Island, NY) containing 10% fetal calf serum (FCS) (Zhejiang Tianhang Biological Technology Co., Ltd., Hangzhou, Zhejiang, China) for Dami cells or fetal bovine serum (FBS) (Hyclone, Logan, UT) for HEL and Meg-01 cells. CMK cells were obtained from the German Collection of Microorganisms and Cell Cultures (DSMZ; Braunschweig, Germany) and cultured in RPMI 1640 medium with 10% FCS. The cultures were maintained in a humidified atmosphere of 5% CO₂ at 37°C. To determine the growth curve, the cells were seeded at 2×10⁵/ml in 10 ml of complete media and cultured in 25-cm² tissue culture flasks (Costar, Corning Incorporated, Corning, NY). Cell counts were performed using the trypan blue dye exclusion method.