Influence of hyperosmolar basal media on hybridoma cell growth and antibody production

Abstract

To investigate the influence of hyperosmolar basal media on hybridoma response, S3H5/gamma 2bA2 and DB9G8 hybridomas were cultivated in a batch mode using hyperosmolar basal media resulting from additional sodium chloride supplementation. The basal media used in this study were IMDM, DMEM, and RPMI 1640, all of which are widely used for hybridoma cell culture. In IMDM, two hybridomas showed different responses to hyperosmotic stress regarding specific MAb productivity (q(MAb)), though they showed similar depression of cell growth in hyperosmolar media. Unlike S3H5/gamma 2bA2 hybridoma, the q(MAb) Of DB9G8 hybridoma was not enhanced significantly around 390 mOsm kg(-1). The variation of basal media influenced DB9G8 hybridoma response to hyperosmotic stress regarding q(MAb) In IMDM, the q(MAb) Of DB9G8 hybridoma was increased by more than 200% when the osmolality increased from 281 to 440 mOsm/kg. However, in RPMI 1640 and DMEM, similar amplitude of osmolality increase resulted in less than 100% increase in q(MAb) The variation of basal media also influenced the cell growth in hyperosmolar medium. Both hybridomas were more tolerant against hyperosmotic stress in DMEM than in IMDM, which was found to be due to the high osmolality of standard DMEM. The osmolalities of standard IMDM and DMEM used for inocula preparation were 281 and 316 mOsm kg(-1), respectively. Thus, when the cells were cultivated at 440 mOsm kg(-1), the cells in IMDM experienced higher osmotic shock than in DMEM. By using the inoculum prepared at 317 mOsm kg(-1) in IMDM, S3H5/gamma 2bA2 cell growth at 440 mOsm kg(-1) in IMDM was comparable to that in DMEM. Taken together, the results obtained from this study show that the selection of basal media is an important factor for MAb production by employing hyperosmotic stress.