The role of p21cip1 in adaptation of CHO cells to suspension and protein-free culture

Abstract

The up-regulation of cyclin-dependent kinase inhibitor p21 has been shown to enhance productivity of monoclonal antibodies and has been linked to various regulatory processes. To identify the potential role of p21 in adaptation to suspension and protein-free cultures, we studied the survival and growth of anchorage- and serum-dependent CHO cell lines that differed only in the period of p21-induced arrest. p21 overexpression led to rapid adaptation of cells to suspension and protein-free cultures. The period taken to achieve adaptation was correlated with the time the cells were arrested after transfer from the monolayer and serum-fed culture. Interestingly, cell aggregation associated with protein-free suspension culture was reduced in p21 culture in response to the loss of cellular adherence. The processes of adaptation to suspension and arrest did not decrease monoclonal antibody productivity. In contrast, following adaptation to protein-free growth media, an overall increase in specific productivity was observed. The ability of cells to survive in protein-free suspension cultures was due to the requirement of G1 cells to growth factors and to their relatively high resistance to the hydrodynamic forces. This improved process has the advantage of reducing the duration of critical path activity for developing CHO commercial cell lines from 72 to 36 days.