LASP-02: In Vitro Cell Cytotoxicity Evaluation in Kras/p53 Pancreatic Ductal Adenocarcinoma (PDAC) Derived Mouse Cells

The Laboratory Animal Sciences Program will assess the in vitro potency of candidate compounds via a conventional cell-based toxicity assay (XTT living cell test) in a series of six drug concentrations (ranging from 0.1 nM to 50,000 nM) of a single agent using at least two independent KPC tumor-derived primary cell lines. In each plate, controls will include dimethyl sulfoxide (DMSO) only (vehicle), 200 nM and 800 nM doxorubicin (dose-dependent response control to chemotherapy agent), gemcitabine, and 0.1% triton (cell-growth negative control). Drug-treated and control assays will be conducted in triplicate. Cell viability will be measured 72 hours after drug exposure using the XTT assay (Roche) per manufacturer’s instruction. Cell viability response to experimental compound will be expressed as a percentage of DMSO (or appropriate vehicle) treated control. The viability of at least one human pancreatic ductal adenocarcinoma (PDAC) cell line (KrasG12D; p53R172H; or p53-/-) will be well tested in response to the compound of interest. 

This service will take about three weeks to complete, and the Laboratory Animal Sciences Program will provide a detailed evaluation (in tabulated and graphical formats) of XTT assay data. Sufficient basic consultation with a subject matter expert is included with the service.

LASP-08: In Vitro Cell Cytotoxicity Evaluation in Kras/p53 Non-Small Cell Lung Cancer Derived Mouse Cells

The Laboratory Animal Sciences Programwillplate primary lung adenocarcinoma cells derived from KrasG12D;p53-/- mice at a routine cell density of 5,000 cells per well in 96-well plate. Cells will be allowed to adhere for 16 hours before the medium will be replaced and the cells will be treated with the candidate compound (AE1). Single drug treatments will be conducted at increasing concentrations from 0.1 nM to 50,000 nM in 0.5% dimethyl sulfoxide (DMSO) unless preliminary compound potency data warrants an increase or decrease in concentration range. Each plate controls will include DMSO only (cell growth in vehicle), 200 nM and 800 nM doxorubicin (known dose-dependent response control), and 0.1% triton (cell growth negative control). All treatments will be run in triplicate. Cell viability will be measured after 72 hours of drug exposure using an XTT assay (Roche) per manufacturer’s instruction. DMSO-treated wells are considered as 100% viability for each treatment plate. Cell viability response to the requestor’s compound will be expressed as a percentage of the DMSO-treated control. Each plate will also include treatment with a standard of care compound (cisplatin or carboplatin) for comparison of response. The entire assay will be repeated in an additional KrasG12D;p53-/- primary tumor line for comparison. 

This service will take about three weeks to complete, and the Laboratory Animal Sciences Program will provide a detailed evaluation (in tabulated and graphical formats) of XTT assay data. Sufficient basic consultation with a subject matter expert is included with the service.