High‐density microcarrier cell cultures for influenza virus production

Böck, A.; Schulze‐Horsel, J.; Schwarzer, J.; Rapp, Erdmann; Genzel, Yvonne; Reichl, Udo

doi:10.1002/btpr.539

Cited by 29 publications

(34 citation statements)

References 44 publications

(55 reference statements)

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“…The cultivation was performed under glucose and glutamine-limited conditions in fed-batch mode and, as a consequence, the cells metabolized these substrates more efficiently (Ljunggren and Häggström, 1994). For highdensity cultivation of adherent MDCK cells a repeated fedbatch operation was established, which also reduced the concentration of unwanted by-products of the cellular metabolism by removal of medium after defined time intervals (Bock et al, 2010). Irani et al (1999) presented a different approach to avoid ammonia accumulation.…”

Section: Introductionmentioning

confidence: 99%

Metabolic adaptation of MDCK cells to different growth conditions: Effects on catalytic activities of central metabolic enzymes

Janke¹,

Genzel²,

Händel³

et al. 2011

Biotech & Bioengineering

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Lactate and ammonia are the most important waste products of central carbon metabolism in mammalian cell cultures. In particular during batch and fed-batch cultivations these toxic by-products are excreted into the medium in large amounts, and not only affect cell viability and productivity but often also prevent growth to high cell densities. The most promising approach to overcome such a metabolic imbalance is the replacement of one or several components in the culture medium. It has been previously shown that pyruvate can be substituted for glutamine in cultures of adherent Madin-Darby canine kidney (MDCK) cells. As a consequence, the cells not only released no ammonia but glucose consumption and lactate production were also reduced significantly. In this work, the impact of media changes on glucose and glutamine metabolism was further elucidated by using a high-throughput platform for enzyme activity measurements of mammalian cells. Adherent MDCK cells were grown to stationary and exponential phase in six-well plates in serum-containing GMEM supplemented with glutamine or pyruvate. A total number of 28 key metabolic enzyme activities of cell extracts were analyzed. The overall activity of the pentose phosphate pathway was up-regulated during exponential cell growth in pyruvate-containing medium suggesting that more glucose-6-phosphate was channeled into the oxidative branch. Furthermore, the anaplerotic enzymes pyruvate carboxylase and pyruvate dehydrogenase showed higher cell specific activities with pyruvate. An increase in cell specific activity was also found for NAD(+)-dependent isocitrate dehydrogenase, glutamate dehydrogenase, and glutamine synthetase in MDCK cells grown with pyruvate. It can be assumed that the increase in enzyme activities was required to compensate for the energy demand and to replenish the glutamine pool. On the other hand, the activities of glutaminolytic enzymes (e.g., alanine and aspartate transaminase) were decreased in cells grown with pyruvate, which seems to be related to a decreased glutamine metabolism.

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Section: Introductionmentioning

confidence: 99%

Metabolic adaptation of MDCK cells to different growth conditions: Effects on catalytic activities of central metabolic enzymes

Janke¹,

Genzel²,

Händel³

et al. 2011

Biotech & Bioengineering

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“…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.…”

Section: Current Strategies To Overcome Nutritional Limitationmentioning

confidence: 99%

“…[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.…”

Section: Introductionmentioning

confidence: 99%

Overcoming nutrient limitations for cell-based production of influenza vaccine

Liu

Ding

Tan

et al. 2015

Human Vaccines & Immunotherapeutics

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“…In recent years, tremendous progress has been made in improving the cultivation of HPAIVs in MDCK cells using microcarrier beads in bioreactors (Bock et al 2011;Genzel et al 2004;Genzel et al 2006;Hu et al 2011;Hu et al 2008;Liu et al 2012;Tree et al 2001), and some investigations on the immunogenicity and reactogenicity of the MDCK-derived HPAIV vaccines have been reported (Doroshenko and Halperin 2009;Palache et al 1997). Nevertheless, differences among the influenza virus subtypes require modifications to the microcarrier-based MDCK cell culture conditions, including the medium formulation, agitation speed, pH, dissolved oxygen, and temperature (Bock et al 2011;Genzel et al 2004;Genzel et al 2006;Hu et al 2011;Hu et al 2008;Liu et al 2012;Tree et al 2001).…”

Section: Introductionmentioning

confidence: 98%

“…Moreover, the emergence of new viral strains responsible for global pandemics requires a switch in vaccine production from the seasonal to the pandemic vaccine. However, the pandemic vaccine manufacturing processes are currently a bottleneck and unable to meet the global vaccination demand (Bock et al 2011;Hu et al 2011;Liu et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Rapid production of a H9N2 influenza vaccine from MDCK cells for protecting chicken against influenza virus infection

Ren

Lu²,

Wang

et al. 2015

Appl Microbiol Biotechnol

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H9N2 subtype avian influenza viruses are widespread in domestic poultry, and vaccination remains the most effective way to protect the chicken population from avian influenza pandemics. Currently, egg-based H9N2 influenza vaccine production has several disadvantages and mammalian MDCK cells are being investigated as candidates for influenza vaccine production. However, little research has been conducted on low pathogenic avian influenza viruses (LPAIV) such as H9N2 replicating in mammalian cells using microcarrier beads in a bioreactor. In this study, we present a systematic analysis of a safe H9N2 influenza vaccine derived from MDCK cells for protecting chickens against influenza virus infection. In 2008, we isolated two novel H9N2 influenza viruses from chickens raised in southern China, and these H9N2 viruses were adapted to MDCK cells. The H9N2 virus was produced in MDCK cells in a scalable bioreactor, purified, inactivated, and investigated for use as a vaccine. The MDCK-derived H9N2 vaccine was able to induce high titers of neutralizing antibodies in chickens of different ages. Histopathological examination, direct immunofluorescence, HI assay, CD4(+)/CD8(+) ratio test, and cytokine evaluation indicated that the MDCK-derived H9N2 vaccine evoked a rapid and effective immune response to protect chickens from influenza infection. High titers of H9N2-specific antibodies were maintained in chickens for 5 months, and the MDCK-derived H9N2 vaccine had no effects on chicken growth. The use of MDCK cells in bioreactors for LPAIV vaccine production is an attractive option to prevent outbreaks of LPAIV in poultry.

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High‐density microcarrier cell cultures for influenza virus production

Cited by 29 publications

References 44 publications

Metabolic adaptation of MDCK cells to different growth conditions: Effects on catalytic activities of central metabolic enzymes

Metabolic adaptation of MDCK cells to different growth conditions: Effects on catalytic activities of central metabolic enzymes

Overcoming nutrient limitations for cell-based production of influenza vaccine

Rapid production of a H9N2 influenza vaccine from MDCK cells for protecting chicken against influenza virus infection

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