High cell density cultivations by alternating tangential flow (ATF) perfusion for influenza A virus production using suspension cells

Overcoming nutrient limitations for cell-based production of influenza vaccine

“…2). 18 However, whether the nutritional supplementation is entirely dedicated to influenza virus production in these studies remains unclear. It is highly possible that not all nutrients consumed by cells enter the pro-viral pathway.…”

“…To address this issue, , current strategies include both simple and straightforward processes of fresh medium addition 12 and medium exchanging, and the relatively complex processes like fedbatch and perfusion. [13][14][15][16][17][18][19] In particular, one reported approach utilizes an alternating tangential flow (ATF) system. 18 With eight successful high cell-density ATF perfusion runs for the production of H1N1 virus, it was demonstrated that cell-specific virus yields could be kept constant and cell-specific infectious virus titers were even higher than those of batch cultures (Fig.…”

“…[9][10][11] Not surprisingly, the common maintaining media used in virus production processes cannot provide sufficient nutrients in general and additional supplementation of nutrients must be performed. Currently, several strategies have been exploited to address this issue of nutritional limitation, including fresh medium replenishment, medium-exchanging, fed-batch, perfusion, etc.. [12][13][14][15][16][17][18] However, a rational design of medium supplementation based on comprehensive nutritional analyses for cells during virus production phase is still missing. Only through a detailed analysis on the differences in the cellular metabolism before and after viral infection, an insight understanding of real nutritional requirements during virus production phase can be achieved, and a rational design of medium supplementation can be solid.…”

Cell culture-based influenza vaccines: A necessary and indispensable investment for the future

“…The cells can be easily adapted to and be grown in serum-free media, and in suspension, as well as on various microcarriers maintained under various bioreactor conditions. 93,[101][102][103][104][105][106][107][108] Subclones of MDCK cells adopted to grow in suspension and support robust virus production have also been described, 91,108,109 although adherent MDCK cells appear to support more robust virus production than suspension MDCK cells. 110 Influenza vaccines derived from MDCK cells are also safe and immunogenic.…”

“…The AGE1.CR cell line was derived by transforming Muscovy duck embryo retinal cells through stable transfection with human adenovirus type 5 E1A/ E1B genes. The cells have been extensively characterized to meet regulatory requirements, and can grow in suspension, although they require the addition of trypsin for influenza virus propagation, 105,[142][143][144] The EB66 cell line was derived as a stable line through a non-chemical non-genetic selection process, and can be adopted to various culture conditions, including suspension culture, bioreactor conditions and to serum-free or chemically defined media, and can grow to high density. 145 Another avian cell line potentially useful for influenza virus production is the immortalized chick embryo cell line PBS-1.…”