Four human colorectal adenocarcinoma tumor cell lines, previously established and characterized in monolayer culture were grown in a matrix-perfusion culture system to determine the suitability of this technique for synthesis of carcinoembryonic antigen (CEA). Production of CEA in excess of 100,000 ng was attained from one cell line, SW 403, during 15-day growth trials. In growth trials and cell-free diffusion studies, CEA passed through membranes of 100,000-dalton molecular weight porosity but not 10,000 porosity. Using cell cultures of high, moderate, or low producers, CEA synthesis tended to reach a plateau after several days of culture and remained nearly constant as the cells attained a maintenance condition. Basic biologic characteristics of the cell lines, expressed as growth rates and CEA produced per 10(6) cells, were comparable in monolayer and perfusion culture. The high cell densities, (10(8) to 10(9) cells per ml) achieved in matrix perfusion made it possible to routinely obtain continuous high yields of CEA over an extended time period.
An ex vivo culture system was developed for assessing the activity of cancer chemotherapeutic agents against tumor cells. The system utilizes artificial capillary culture units and the technique of hemodialysis to expose tumor cells to a chemotherapeutic drug and its metabolites following injection of the drug into an experimental animal. This ex vivo culture system was used to test the activity of 5-fluorouracil (5-FU) against four human colorectal adenocarcinoma cell lines (SW 403, SW 480, SW 620, and SW 707). Cell killing by 5-FU or its metabolites in blood dialysate following intravenous injection was measured by determining colony formation of cells attached to plastic and suspended in 0.3% agar after short-term exposures of 1 to 2 h. The technique was shown to discriminate between the sensitivities of these cell lines and the respective sensitivities to the drug were reproducible. Kinetics of drug clearance from the host animal's blood were shown to be similar to that in humans. The results suggest the system may be useful for testing diverse drugs, including those requiring metabolic activation, against a variety of types of tumor cells.
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