Cell mediated lympholysis: CML. a microplate technique requiring few target cells and employing a new method of supernatant collection

Abstract

A micro method for the 51Cr release assay is described. Allogeneically induced cytotoxic lymphocytes are generated in micro mixed lymphocyte cultures in the wells of micro plates. Their cytotoxic capacity is assayed by adding 51Cr labelled PHA derived lymphoblasts directly into the micro cultures with no pooling or transfer of the cytotoxic effector cells being required. The 51Cr isotope released into the cell supernatants is collected by inserting a cellulose acetate absorption cartridge into each well. A glass fiber filter attached to the cartridge effectively separates the supernatant from the cellular elements. This system allows the simultaneous collection of the supernatant from 96 wells, and can be used with either adherent or non-adherent target cells.