Immune Responses in Vitro

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Abstract
At various intervals after initiation of Mishell-Dutton spleen cell cultures, cell clusters were separated from non-clustered, single spleen cells by sedimentation through a fetal calf serum gradient at unit gravity. Cell clusters collected at all intervals during the initial 48 hr of culture failed to develop subsequently significant plaque-forming cell (PFC) responses. In contrast, the single, non-clustered spleen cells isolated at these intervals subsequently developed PFC responses equal to or greater than responses in unseparated control cultures. Sequential removal of the cell clusters which reformed when the single spleen cells were returned to culture had little or no effect on the PFC responses generated by the remaining single cells. Thus, it appears that the continuous integrity of cell clusters which form in mouse spleen cell cultures has no direct effect on the immune responses which develop in these cultures.

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