Cytotoxicity Screening
All compounds deposited in the IOCB library undergo basic cytotoxicity screening automatically. The initial screening concentration is 10 µM. Dose-response curves are generated for all active compounds, i.e. inducing 50% decrease in cell viability. The tabular data (% viability at 10 µM and IC50 values) are inserted in the database (AHL) where they remain visible to the individual research group members. For installation of AHL database, contact Pavel Šácha.)
Dose-response curves and RAW data are available upon request.
Basic panel of cell lines (all human):
- CCRF-CEM – T-lymphoblastic leukemia (suspension)
- HL-60 – promyelocytic leukemia (suspension)
- HepG2 – hepatocellular carcinoma (adherent)
- HeLa S3 – HPV-positive cervical carcinoma (adherent)
- NHDF – normal dermal fibroblasts (adherent)
Methodology
Routinely, cytotoxicity is evaluated upon 72h-exposure of the cells with the compounds. The detection method is based on quantification of cellular ATP which correlates with the number of viable cells. To this end, we employ commercial reagent CellTiter-Glo® 2.0 (Promega) with luminescent readout. The tests are conducted in 384-well plates.
Specific requirements
We understand that in specific cases, standard conditions of the test do not match well with your research needs (cell lines, test concentration, time, detection …). Or, you do not have enough compound to be put into the library, or you need to speed up the process significantly to recieve timely feedback (typically in active drug-discovery projects). All your concerns and needs can be negotiated on an individual basis.
Kontakty
- Eva Tloušťová (routine screening, sample submission, data interpretation), ☎ 271/403
- Helena Mertlíková Kaiserová (specific requirements, implementation of cytotoxicity screening in drug discovery screening loops), ☎ 114