Antigen Expression |
HLA-A02+, MHC class I - +MHC class II -, , ICAM-1 + ?-microglobulin +, Beta-2-M+, HLA-E+, HLA-G low, , MIC A low, MIC-B -, GFAP+ , nestin +, vimentin +, S-100+, GBM+, BTSC+ |
Mutational Profile |
IDH 1 2 wt, TP53wt, K-Ras G12D, B-RAFwt , 4q12(PDGFRA) amplified, PTEN P169S/del 212-229 |
Subculturing |
Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add TrypLE Express (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at 37 ?C for 10 to 15 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300xg, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. This cell line will result in single cell suspension. |