A simple method of cell-mediated cytotoxic assay by postlabeling of tritiated thymidine.

Abstract

The cytotoxic reaction of effector cells on target cells was performed on cover slips which were set in a petri dish so as to be interposed from the dish surface through the bases of the slide glasses. After incubation, the target cells remaining on the cover slips were pulse-labeled with 3H-TdR for 45 min. Subsequently, the cover slips were removed from the bases, and dipped in cold 5% TCA to be freed from non-incorporated isotopes. The residual target cells adhering on the cover slips were counted without being removed.By this assay system, the specific or non-specific cytotoxicity of lymphocytes or macrophages was easily demonstrated with reliability. -cell-mediated cytotoxic assay; 3H -thymidine; cover slip; lymphocyte; macrophage

Inhibition of Cellular DNA Synthesis by Rat Peritoneal Macrophages

“…Previously, we described that the peritoneal macrophages of the normal rat inhibited DNA synthesis in syngeneic tumor cells induced by 3-methylcholanthrene (Toh and Kikuchi 1977). In this report, additional studies were undertaken to elucidate the regulatory function of macrophages on cellular DNA synthesis.…”

Measurement of Cell Mediated Cytotoxicity by Post‐Labeling Surviving Target Cells

“…Other assays, however, which could be used for the measurement of cell mediated cytotoxicity on non-adherent target cells are not available. The postlabeling techniques described by Brooks et al (1978), Seeger et al (1974) and Toh & Kikuchi (1977) require the separation of the effector cells from the target cells. This paper describes a new assay where effector cells have not been separated from target cells.…”