Abstract
Previous investigations have shown that a substantial fraction of mouse hybridoma cells in suspension culture die by apoptosis. This work represents a first attempt at timing the onset of spontaneous apoptosis in hybridoma batch culture. Markers of apoptosis were followed in batch cultures carried out in protein-free medium. A steep increase of the apoptotic index, i.e. the relative number of bodies insoluble in 6M guanidinium hydrochloride, was observed on day. A notable increase of the degree of DNA fragmentation, that represents an early event of the death program, was apparent already on day 4. Thus, the critical period of the spontaneous onset of apoptosis is the exponential phase of the culture. This finding indicates that spontaneous apoptosis in mouse hybridomas is a process associated with cell proliferation and full metabolic activity.
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