WHO Requirements for the Use of Animal Cells asin vitroSubstrates for the Production of Biologicals (Requirements for Biological Susbstances No. 50)

Production and characterization of high-titer serum-free cell culture grown hepatitis C virus particles of genotype 1–6

“…These unique characteristics, as well as the reduced concentration of non-HCV proteins in serum-free culture supernatants, are expected to facilitate generation of purified and concentrated virus stocks, required for vaccine development and biophysical studies of HCV particle composition [54–57]. Further, the developed serum-free culture conditions might reduce the risk of contamination with adventitious microbial agents in future vaccine antigen preparations [40,41]. …”

“…Due to their increased permissiveness to infection with recombinant HCV, Huh7.5 cells were previously used for cell culture adaptation and growth of HCV genotype 1–6 recombinants used in this study [11,12,32,34,35]. According to WHO recommendations, a wide range of continuous cell lines are now considered as suitable substrates for production of various medicinal substances if certain requirements are met [40]. These requirements include use of well-characterized cell banks, use of suitable manufacturing procedures aiming at a high degree of purification of the end product, and thorough characterization of the end product [40].…”

“…According to WHO recommendations, a wide range of continuous cell lines are now considered as suitable substrates for production of various medicinal substances if certain requirements are met [40]. These requirements include use of well-characterized cell banks, use of suitable manufacturing procedures aiming at a high degree of purification of the end product, and thorough characterization of the end product [40]. Thus, Huh7.5 cells could potentially be characterized to comply with these recommendations, allowing their use for vaccine development.…”

“…Further, as patient derived HCV particles [17], HCVcc showed a heterogeneous density profile [6,32,38,39], making density-based purification and concentration procedures difficult. Also, cell cultures are typically treated with animal-derived trypsin, and growth medium used for production of HCVcc is typically supplemented with fetal bovine serum (FBS) [40]. Vaccine development, as well as other research applications, such as biophysical studies of HCV particle composition, require generation of purified and concentrated HCVcc stocks.…”

“…This is expected to be facilitated by reducing concentrations of non-HCV proteins such as FBS derived proteins in HCVcc producing cell cultures. Further, use of FBS and animal-derived trypsin increases the risk of contamination with adventitious microbial agents, of relevance for HCV vaccine development [40,41]. Thus, development of methods for production of HCVcc under serum-free conditions is a research focus.…”