Preparative Electrophoretic Separation of Antibody Forming Cells

Abstract

A procedure is described for preparative electrophoretic separation of lymphoid cells. The separations were performed with a free flow electrophoretic cell separator model VAP IV (Desaga, Heidelberg, Bender & Hobein, Munich, Brinkmann Instruments, Westbury, N. Y.). Rats were immunized with sheep erythrocytes (SRBC), lymph node cells electrophoretically separated at different times after immunization and the fractions obtained subsequently cultured in diffusion chambers. The antibody forming cells and the morphological composition of the fractions was determined after separation and after culture. Lymph node cells could be separated into 16 fractions. Within this heterogeneous distribution profile two narrow distributions of antibody forming cells of the same specificity but of different stages of development could be detected. The distribution of lower electrophoretic mobility contained the primed lymphocytes and blast cells, the faster distribution contained the differentiated plasma cells. It was found that a homogeneous cell population is rather homogeneous in its electrophoretic mobility. This is an indication that the electrophoretic mobility would be a useful parameter enabling separation of functionally defined cell populations.