Cancer Cell Culturing and Cytotoxicity Assays for Anticancer Screening at
City of Hope
Research: Response and Resistance of cancer cells to therapeutic agents
Failure of cancer treatment
One main cause: drug resistance by tumor
Breast cancer is the most common cancer in women (incurable if metastatic)
Her2 is overexpressed in 25-30% of breast cancer and helps the cancer resist conventional treatments

Some antibiotics are also anti-cancer drugs
Immune defense in infectious agents and tumor defense display some similarities
Antimicrobial lipids produced naturally in body fluids might exert some anticancer activity

Collaborative Design
Cell Maintenance and Technique
Cell growth and passage
Tissue culture medium
Trypsin, serum, L-glutamine
Cryopreservation and storage
Cell Counting
Coulter counter
Safety and technique

Measuring Cytotoxicity
Clonogenic Assay (colony forming assay)
Sulforhodamine B (SRB Assay)

Colony Forming Assay
Treatment is applied to a sample of cells.
The cells are plated in a tissue culture vessel and allowed to grow.
The colonies produced are fixed, stained, and counted
Cell survival – surviving fraction vs. drug concentration

Slide 8
The sulforhodamine B (SRB) Assay
SRB binds to protein components of cell
SRB is bright-pink aminoxanthene dye and binds basic amino groups – binding is stoichiometric to cell mass
Adaptable to 96-well, high throughput screening
Dose Response Analysis

Herceptin Study
Herceptin (trastuzumab) – monoclonal antibody
Used to treat breast cancers over-expressing Her2/neu receptor
Disrupt cell signalling pathway –
PI-3-kinase/Akt signal transduction pathway

First Results
Next Steps
Complete training at City of Hope
Implement various screening protocols with antimicrobial lipid based liposome preparations privided by Molecular Express, Inc. at CSULA