Abstract

Animal cell culture, with single cells growing in suspension, ideally in a chemically defined environment, is a mainstay of biopharmaceutical production. The synthetic environment lacks exogenous growth factors and usually requires a time-consuming adaptation process to select cell clones that proliferate in suspension to high cell numbers. The molecular mechanisms that facilitate the adaptation and that take place inside the cell are largely unknown. Especially for cell lines that are used for virus antigen production such as baby hamster kidney (BHK) cells, the restriction of virus growth through the evolution of undesired cell characteristics is highly unwanted. The comparison between adherently growing BHK cells and suspension cells with different susceptibility to foot-and-mouth disease virus revealed differences in the expression of cellular receptors such as integrins and heparan sulfates, and in the organization of the actin cytoskeleton. Transcriptome analyses and growth kinetics demonstrated the diversity of BHK cell lines and confirmed the importance of well-characterized parental cell clones and mindful screening to make sure that essential cellular features do not get lost during adaptation.

Highlights

  • Animal cell culture technology was first used for the production of virus antigen in vaccine production, but more recently bacterial or yeast production platforms for recombinant proteins are changed over to mammalian cell systems as well [1, 2]

  • The following cell lines were used in addition: baby hamster kidney (BHK)-2P maintained either in Glasgow’s Minimal Essential Medium (GMEM) with 10% foetal bovine serum (FBS) (“line #2”) or adapted to the Animal-component-free media (ACFM) CellventoTM BHK200 (Merck KGaA, Darmstadt, Germany) in two different processes, plus four other suspension cell lines: BHK21C13 adapted to growth in suspension by gradual withdrawal of serum, BHK21-C, BHK21-Hektor and production BHK [12]

  • Batch experiments with nine different BHK suspension cell lines were conducted over a period of six days and viable cell density (VCD) and viability were measured daily

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Summary

Introduction

Animal cell culture technology was first used for the production of virus antigen in vaccine production, but more recently bacterial or yeast production platforms for recombinant proteins are changed over to mammalian cell systems as well [1, 2]. Important in both contexts are baby hamster kidney (BHK) cells, derived from the Syrian golden hamster (Mesocricetus auratus). BHK cells were an adherently growing, anchor-dependent mammalian cell line with a fibroblast growth pattern in standard cell culture conditions including foetal bovine serum (FBS) [3, 4].

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