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The productivity of recombinant mammalian cell lines growth in batch culture is often limited by the rapidity with which cells die on entry into the decline phase (the period of culture after the maximum cell density has been reached and where cell viability begins to fall). We examined the decline phase characteristics of the NSO myeloma cell line with a view to modulating the cell death that ensues. Examination of nuclear morphology during culture revealed that the onset of the decline phase was marked by a time-dependent increase in the percentage of cells that exhibited condensed and fragmented nuclei. Furthermore, these changes coincided with a fall in DNA integrity. High molecular weight DNA appeared to be degraded into oligonucleosomal fragments. Taken together, these observations indicated that NSO cells die by the process of apoptosis. The protein encoded by the bcl-2 gene has been shown to counter apoptosis induced by a large variety of stimuli and in a number of different cell types, but is not expressed in NSO cells. We examined whether overexpression of this protein could prevent/delay the onset of cell death seen during batch culture and also in response to serum limitation. Bcl-2 failed to affect the decline phase characteristics and serum dependence of NSO cells. In our search to explain these findings, we found that the NSO cell line expresses bax and also a high level of another Bcl-2 related protein, Bcl-x(L). Given that Bcl-X(L) is a sequence and functional homologue of Bcl-2, it is possible that Bcl-2 is redundant in the NSO cell background. These data therefore indicate that cells such as NSO, which are used in biotechnologically important processes such as generation of hybridomas and expression of recombinant proteins, may express only a subset of genes important in apoptotic regulation. Modulation of the death characteristics of such cells will need to take account of the expression profile of such genes and their regulatory interactions.

Original publication




Journal article


Biotechnol Bioeng

Publication Date





298 - 304