INSITU LYMPHOID-CELLS OF MOUSE MAMMARY-TUMORS .1. DEVELOPMENT AND EVALUATION OF A METHOD FOR SEPARATION OF LYMPHOID-CELLS FROM MOUSE MAMMARY-TUMORS

Abstract

A method of separating lymphoid cells from solid mouse mammary tumors was developed and evaluated. In this method the tumors are digested with 0.01% collagenase, 0.0 DNAase and 0.025% trypsin in Dulbecco''s PBS [phosphate buffered saline] into suspensions of cells with a viability of 90%. The suspensions are fractionated on a continuous gradient of Ficoll in tissue culture medium. In model experiments this gradient separates, cleanly, admixed cells of an established mammary tumor cell line and dissociated thymus glands. Recovery rates were 50% for the tumor cells and 80% for the thymocytes. The preparation of the cell suspensions and the gradient separation procedure are not harmful to the cells as indicated by trypan blue exclusion and the ability to grow in cell culture.