Iomm lee

IMR90, normal human fetal lung fibroblasts, and IOMM-Lee, a human malignant meningioma cell line expressing wild-type p53, were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% or 5% fetal bovine serum (FBS), respectively. HKBMM, a human malignant meningioma cell line expressing a mutant p53 (P177L), was obtained from the Riken BioResource Research Center (Tsukuba, Japan) and maintained in Ham’s F12 medium supplemented with 10% FBS. The culture media were supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin. The p53 status of IOMM-Lee and HKBMM cells was examined by cDNA sequencing as described below. IMR90 cells passaged fewer than 8 times were used in experiments. Detailed conditions for the in vitro drug treatment are described in the respective figure legends. In general, the day after cells had been passaged, the required amount of the drug stock solution was added to cells and gently mixed well. This was followed by an incubation at 37 °C in a 5% CO₂ incubator for the specified time, and cells were then subjected to each assay.

Meningioma tumor samples were obtained intraoperatively from patients for whom tumor-banking consent were obtained. All experiments were performed in accordance with our institutional Research Ethics Board at the University Health Network (Toronto, Canada) and the University of Toronto. Cell suspensions were created and maintained as previously reported on ThermoFisher BioLite 100 mm Tissue Culture dishes in DMEM/F12 supplemented with 2 × non-essential amino acids (NEAA), 10 μg/mL gentamicin (Sigma-Aldrich) and fetal bovine serum (10% v/v; Life Technologies, Carlsbad, CA, USA) in an incubator at 37 °C and 5% CO₂. Once confluent, cells were passaged following trypsinization [28 (link), 30 (link)]. The following primary meningioma cell lines were used for in vitro drug screening: mng_20, mng_50, mng_84, mng_46. CH157 (CH-157MN; RRID:CVCL_5723) and IOMM-Lee (RRID:CVCL_5779) immortalized meningioma cell lines were obtained from the American Type Culture Collection (ATCC) and were cultured as monolayers in the same media composition as above.

IOMM-Lee and HKBMM, human malignant meningioma cell lines, were obtained from the American Type Culture Collection (Manassas, VA, USA) and from the Riken BioResource Center (Tsukuba, Japan), respectively. Their characteristics as malignant meningioma cells were confirmed in a previous study [8 (link)]. IOMM-Lee cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum (FBS). HKBMM cells were cultured in Ham’s F12 medium supplemented with 10% FBS.

IOMM-Lee, a human malignant meningioma cell line, and IMR90, normal human fetal lung fibroblasts, were obtained from the American Type Culture Collection (Manassas, VA, USA) and cultured in DMEM supplemented with 5% or 10% fetal bovine serum (FBS), respectively. The human glioblastoma cell lines, A172 and T98G, were provided by the RIKEN RBC (Tsukuba, Japan) through the National BioResource Project of the Ministry of Education, Culture, Sports, Science and Technology, Japan and cultured in RPMI 1640 medium supplemented with 10% FBS. The culture media were supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin. All the IMR90 experiments were performed using cells with a low passage number (<8).