Chemo-sensitivity assays using tumor cells exhibiting persistent phenotypic characteristics

A method for testing sensitivity of a tumor to a therapeutic agent, the method comprising:

• mincing an epithelial tumor specimen to prepare a plurality of multicellular tissue explants, and placing the explants in liquid medium in one or more containers;
• physically agitating the explants, culturing said explants to form one or more non-confluent monolayers, and suspending the non-confluent monolayer(s) in culture medium to obtain from about 4,000 to about 12,000 cells per ml;
• transferring the cells to a plurality of wells at about 100 to about 1000 cells per well;
• adding a serial dilution of at least one therapeutic agent or combination across the wells, wherein the therapeutic agent comprises a monoclonal antibody;
• and
• determining sensitivity of the cells to the therapeutic agent.

A method for testing sensitivity of a tumor to a therapeutic agent, the method comprising:

• disaggregating an ovarian or colorectal tumor specimen to prepare a plurality of explants;
• treating the explants with DNase and Collagenase while disturbing the explants with a swirling, shaking, or rocking motion so as to reduce the size of the explants, wherein the treated explants are multicellular tissue explants;
• culturing the multicellular tissue explants to form one or more non-confluent monolayers comprising malignant cells;
• suspending the non-confluent monolayer(s) in culture medium at about 4,000 to 12,000 cells per ml, and transferring the cells to a plurality of wells at about 100 to 1000 cells per well;
• incubating the cells for about 4 to 30 hours, and then adding a serial dilution of the therapeutic agent across the wells, wherein the therapeutic agent comprises a monoclonal antibody;
• quantifying cell death in each well as a measure of sensitivity;
• and
• preparing a dose response curve for the therapeutic agent.