Plaque Forming Cells Produced in Test Tube Cultures of Normal Mouse Spleen Cells Incubated with Erythrocytes 1

Summary

Test tube cultures of normal mouse spleen cells set up as for mixed leukocyte culture, but with antigenic RBC, and fed once with additional medium without disturbing the button of cells, were found to contain antibody producing cells, as indicated by the ability of such cells to produce hemolytic antibody plaques when plated against the RBC. In the case of sheep RBC as antigen, the number of PFC increased to day 7, when a level of 90-110 per 10⁶ starting spleen cells was reached, in comparison with 18-24 PFC in control cultures, without antigen. With burro RBC such results were also obtained, with somewhat lower levels of PFC produced in antigen-containing cultures, but substantially lower background levels. Since dispersion of the cells at re-feeding caused failure of appearance of the PFC, a condition for the production of these was presumably the formation of a microenvironment within the button of cells in the bottom of the tube.