Wave microcarrier cultivation of MDCK cells for influenza virus production in serum containing and serum-free media

Genzel, Yvonne; Olmer, Ruth; Schäfer, Bernd; Reichl, Udo

doi:10.1016/j.vaccine.2006.05.023

Cited by 95 publications

(67 citation statements)

References 29 publications

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“…would be equivalent to about 30 eggs. In another study, titer of 2.5 ϫ 10 3 HAU/mL was obtained in a stirred tank reactor using Cytodex I microcarriers (16) and a maximum titer of approximately 4.0 ϫ 10 4 HAU/mL was obtained in WAVE cellbags using Cytodex I microcarriers (18). It is important to note that in these studies the Influenza A virus strain was used.…”

Section: Discussionmentioning

confidence: 99%

“…Without surface attachment, these cells cannot exert their normal cyclindependent kinase activity through the signaling cascades initialized by interactions between integrins and extracellular matrix (11)(12)(13)(14)(15). For industrial production in bioreactors, the required surface area can be provided using microcarrier beads (16)(17)(18)(19). Although this approach is sufficient to obtain high virus production yield (18,19), this propagation strategy is cumbersome compared with propagation of cells in suspension.…”

mentioning

confidence: 99%

“…For industrial production in bioreactors, the required surface area can be provided using microcarrier beads (16)(17)(18)(19). Although this approach is sufficient to obtain high virus production yield (18,19), this propagation strategy is cumbersome compared with propagation of cells in suspension. An MDCK cell line that can proliferate in suspension would greatly facilitate the scale-up process of influenza virus production.…”

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confidence: 99%

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Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

Chu

Lugovtsev

Golding

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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MDCK cells are currently being considered as an alternative to embryonated eggs for influenza virus propagation and hemagglutinin (HA) production intended for vaccine manufacturing. MDCK cells were found suitable for the virus production but their inability to grow in suspension burdens the process of scale up and hence their production capability. Anchorage-dependent MDCK cells were converted to anchorage-independent cells, capable of growing in suspension as a result of transfection with the human siat7e gene (ST6GalNac V). This gene was previously identified as having an important role in cellular adhesion when the transcriptions of genes from anchorage-dependent and anchorage-independent HeLa cells were compared. Unlike the parental MDCK cells, the siat7e-expressing cells were capable of growing in shake flasks as suspension cultures, achieving maximum concentration of 7 ؋ 10 5 cells/mL while keeping close to 100% viability throughout the growth phase. In production experiments, the siat7e-expressing cells were infected with the Influenza B/Victoria/504/2000 strain. It was determined that the cell-derived viruses retained similar antigenic properties as those obtained from egg-derived viruses and their nucleotide sequences were identical. The specific production of hemagglutinin (expressed in hemagglutination units per 10 6 cells) from the siat7e-expressing cells was approximately 20 times higher than the specific production from the parental MDCK cells. If this suspension process scales up, the production potential of HA from 10 L of siat7e-expressing cells at a concentration of 10 6 cells/mL would be equivalent to the amount of HA obtained from 10,000 embryonated eggs.anchorage-independent ͉ hemagglutinin ͉ sialyltransferase ͉ vaccine

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

Chu

Lugovtsev

Golding

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…Hence, the nutritional demand of host cells should undergo a profound and complex shift. For instance, the glucose demand can be affected by virus infection and replication, [4][5][6] various proteins need to be synthesized for virus replication, which apparently exert significant influences on the demand for amino acids, 7,8 and lipid rafts are also badly needed during the infection and replication process. [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.…”

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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“…Single use disposable bioprocessing systems for animal cell cultivation have seen a fast evolution in the last few years (Genzel et al, 2006). Single Use Bioreactor (SUB; Thermo Scientific Hyclone 1 , Logan, UT) is a disposable alternative to conventional stirred tank reactors (STR) that are widely used for animal cell culture.…”

Section: Introductionmentioning

confidence: 99%

Production of cell culture (MDCK) derived live attenuated influenza vaccine (LAIV) in a fully disposable platform process

George

Farooq

Dang

et al. 2010

Biotech & Bioengineering

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The majority of influenza vaccines are manufactured using embryonated hens' eggs. The potential occurrence of a pandemic outbreak of avian influenza might reduce or even eliminate the supply of eggs, leaving the human population at risk. Also, the egg-based production technology is intrinsically cumbersome and not easily scalable to provide a rapid worldwide supply of vaccine. In this communication, the production of a cell culture (Madin-Darby canine kidney (MDCK)) derived live attenuated influenza vaccine (LAIV) in a fully disposable platform process using a novel Single Use Bioreactor (SUB) is presented. The cell culture and virus infection was maintained in a disposable stirred tank reactor with PID control of pH, DO, agitation, and temperature, similar to traditional glass or stainless steel bioreactors. The application of this technology was tested using MDCK cells grown on microcarriers in proprietary serum free medium and infection with 2006/2007 seasonal LAIV strains at 25-30 L scale. The MDCK cell growth was optimal at the agitation rate of 100 rpm. Optimization of this parameter allowed the cells to grow at a rate similar to that achieved in the conventional 3 L glass stirred tank bioreactors. Influenza vaccine virus strains, A/New Caledonia/20/99 (H1N1 strain), A/Wisconsin/67/05 (H3N2 strain), and B/Malaysia/2506/04 (B strain) were all successfully produced in SUB with peak virus titers > or =8.6 log(10) FFU/mL. This result demonstrated that more than 1 million doses of vaccine can be produced through one single run of a small bioreactor at the scale of 30 L and thus provided an alternative to the current vaccine production platform with fast turn-around and low upfront facility investment, features that are particularly useful for emerging and developing countries and clinical trial material production.

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Wave microcarrier cultivation of MDCK cells for influenza virus production in serum containing and serum-free media

Cited by 95 publications

References 29 publications

Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production

Overcoming nutrient limitations for cell-based production of influenza vaccine

Production of cell culture (MDCK) derived live attenuated influenza vaccine (LAIV) in a fully disposable platform process

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