ChemoFx® Laboratory Methodology

Specimens are received by Precision's laboratory in shipping media provided to clients by Precision. Shipping studies indicate that tumor integrity is normally maintained during transit times in accordance with Precision's shipping instructions.

Processing specimens

   • Once the tumor specimen arrives at the laboratory, it is removed from the shipping media
     vial and is processed for culture.
   • Any necrotic material or adherent normal tissue is removed from the specimen.
   • The remaining tumor specimen is aseptically minced into small pieces. These pieces are
     transferred to incubation flasks, appropriate culture media is added, and primary explant
     cultures are established.
   • The primary explant cultures are monitored for growth.
   • Once a sufficient number of cells have grown in culture, the cells are trypsinized and
     counted, so that a specific number of cells can be delivered to each well of a
     microtiter plate.
   • After overnight incubation to allow adherence to the bottom of the wells of the microtiter
     plate, cells are then exposed to multiple doses of chemotherapeutic agents. Length of drug
     exposure is based on type of drug selected.
   • Following acute drug exposure, the cells are rinsed, fixed, and stained, therby removing the
     dead, non-adherent cells and staining the remaining live cells with a fluorescent
     nuclear stain.
   • Surviving cell numbers are counted using an automated microscopic image analysis
     system and the fraction surviving is calculated for each drug and dose level.
   • Results are compared to control levels and the data is summarized and
     reported graphically.
   • From these data, a dose-response graph is derived that provides the physician with
     information on the patient's response to each specified anticancer agent.