Production of reovirus type-1 and type-3 from Vero cells grown on solid and macroporous microcarriers

Two strains of reovirus were propagated in Vero cells grown in stationary or microcarriers cultures. Vero cells grown as monolayers on T‐flasks or in spinner cultures of Cytodex‐1 or Cultispher‐G microcarriers could be infected with reovirus serotype 1, strain Lang (T1L), and serotype 3, strain Dear...

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Veröffentlicht in:Biotechnology and bioengineering 1999-01, Vol.62 (1), p.12-19
Hauptverfasser: Berry, J. M., Barnabé, N., Coombs, K. M., Butler, M.
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Butler, M.
description Two strains of reovirus were propagated in Vero cells grown in stationary or microcarriers cultures. Vero cells grown as monolayers on T‐flasks or in spinner cultures of Cytodex‐1 or Cultispher‐G microcarriers could be infected with reovirus serotype 1, strain Lang (T1L), and serotype 3, strain Dearing (T3D). A regime of intermittent low speed stirring at reduced culture volume was critical to ensure viral infection of cells in microcarrier cultures. The virus titre increased by 3 to 4 orders of magnitude over a culture period of 150 h. Titres of the T3D reovirus strain were higher (43%) compared to those of the T1L strain in all cultures. Titres were significantly higher in T‐flask and Cytodex‐1 microcarrier cultures compared to Cultispher‐G cultures with respect to either reovirus type. The viral productivity in the microcarrier cultures was dependent upon the multiplicity of infection (MOI) and the cell/bead ratio at the point of infection. A combination of high MOI (5 pfu/cell) and high cell/bead loading (>400 for Cytodex‐1 and >1,000 for Cultispher‐G) resulted in a low virus productivity per cell. However, at low MOI (0.5 pfu/cell) the virus productivity per cell was significantly higher at high cell/bead loading in cultures of either microcarrier type. The maximum virus titre (8.5 × 109 pfu/mL) was obtained in Cytodex‐1 cultures with a low MOI (0.5 pfu/cell) and a cell/bead loading of 1,000. The virus productivity per cell in these cultures was 4,000 pfu/cell. The lower viral yield in the Cultispher‐G microcarrier cultures is attributed to a decreased accessibility of the entrapped cells to viral infection. The high viral productivity from the Vero cells in Cytodex‐1 cultures suggests that this is a suitable system for the development of a vaccine production system for the Reoviridae viruses. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 62: 12–19, 1999.
doi_str_mv 10.1002/(SICI)1097-0290(19990105)62:1<12::AID-BIT2>3.0.CO;2-G
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M. ; Barnabé, N. ; Coombs, K. M. ; Butler, M.</creator><creatorcontrib>Berry, J. M. ; Barnabé, N. ; Coombs, K. M. ; Butler, M.</creatorcontrib><description>Two strains of reovirus were propagated in Vero cells grown in stationary or microcarriers cultures. Vero cells grown as monolayers on T‐flasks or in spinner cultures of Cytodex‐1 or Cultispher‐G microcarriers could be infected with reovirus serotype 1, strain Lang (T1L), and serotype 3, strain Dearing (T3D). A regime of intermittent low speed stirring at reduced culture volume was critical to ensure viral infection of cells in microcarrier cultures. The virus titre increased by 3 to 4 orders of magnitude over a culture period of 150 h. Titres of the T3D reovirus strain were higher (43%) compared to those of the T1L strain in all cultures. Titres were significantly higher in T‐flask and Cytodex‐1 microcarrier cultures compared to Cultispher‐G cultures with respect to either reovirus type. The viral productivity in the microcarrier cultures was dependent upon the multiplicity of infection (MOI) and the cell/bead ratio at the point of infection. A combination of high MOI (5 pfu/cell) and high cell/bead loading (&gt;400 for Cytodex‐1 and &gt;1,000 for Cultispher‐G) resulted in a low virus productivity per cell. However, at low MOI (0.5 pfu/cell) the virus productivity per cell was significantly higher at high cell/bead loading in cultures of either microcarrier type. The maximum virus titre (8.5 × 109 pfu/mL) was obtained in Cytodex‐1 cultures with a low MOI (0.5 pfu/cell) and a cell/bead loading of 1,000. The virus productivity per cell in these cultures was 4,000 pfu/cell. The lower viral yield in the Cultispher‐G microcarrier cultures is attributed to a decreased accessibility of the entrapped cells to viral infection. 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M.</creatorcontrib><creatorcontrib>Barnabé, N.</creatorcontrib><creatorcontrib>Coombs, K. M.</creatorcontrib><creatorcontrib>Butler, M.</creatorcontrib><title>Production of reovirus type-1 and type-3 from Vero cells grown on solid and macroporous microcarriers</title><title>Biotechnology and bioengineering</title><addtitle>Biotechnol. Bioeng</addtitle><description>Two strains of reovirus were propagated in Vero cells grown in stationary or microcarriers cultures. Vero cells grown as monolayers on T‐flasks or in spinner cultures of Cytodex‐1 or Cultispher‐G microcarriers could be infected with reovirus serotype 1, strain Lang (T1L), and serotype 3, strain Dearing (T3D). A regime of intermittent low speed stirring at reduced culture volume was critical to ensure viral infection of cells in microcarrier cultures. The virus titre increased by 3 to 4 orders of magnitude over a culture period of 150 h. 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The lower viral yield in the Cultispher‐G microcarrier cultures is attributed to a decreased accessibility of the entrapped cells to viral infection. The high viral productivity from the Vero cells in Cytodex‐1 cultures suggests that this is a suitable system for the development of a vaccine production system for the Reoviridae viruses. © 1999 John Wiley &amp; Sons, Inc. Biotechnol Bioeng 62: 12–19, 1999.</description><subject>Animal cells</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Cercopithecus aethiops</subject><subject>Culture Media</subject><subject>Establishment of new cell lines, improvement of cultural methods, mass cultures</subject><subject>Eukaryotic cell cultures</subject><subject>Evaluation Studies as Topic</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Growth kinetics</subject><subject>macroporous</subject><subject>Mammalian orthoreovirus 3 - physiology</subject><subject>Methods. Procedures. Technologies</subject><subject>Microbiology</subject><subject>microcarrier</subject><subject>Microspheres</subject><subject>Orthoreovirus - physiology</subject><subject>reovirus</subject><subject>Reovirus 1</subject><subject>Reovirus 2</subject><subject>Vero</subject><subject>Vero Cells</subject><subject>Viral Plaque Assay</subject><subject>Viral Vaccines - isolation &amp; purification</subject><subject>Virus Cultivation - methods</subject><subject>Virus Replication</subject><subject>Viruses</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1999</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkV1v0zAUhiMEYmXwF1AuENouUnyc2I7Lh7Rl0BZNKxIDLo8cx0GBJC52y-i_x1lLmQRSr-wjvef1Yz1R9BrIGAihL04-zov5KRApEkIlOQEpJQHCTjmdwCugk8nZ_CI5n1_TN-mYjIvFS5pM70Wj_cb9aEQI4UnKJD2KHnn_LYwi5_xhdBRekJKRfBSZD85Wa71qbB_bOnbG_mzc2serzdIkEKu-2l7TuHa2iz8bZ2Nt2tbHX529CTt97G3bVLfJTmlnl9bZUNA14a6Vc41x_nH0oFatN09253H06d3b62KWXC6m8-LsMtGMSprkuoTaEEg5VEprAkIILVheViVTVZjBpNSUWcaJ0WDKStIsJ0ZAxcKiKNPj6Pm2d-nsj7XxK-waP-Cq3gQo5JJzkcvsYJAGhpxJdjAIAjLJgKb7L4Vve-9MjUvXdMptEAgORhEHozj4wcEP_jGKnCIghEAwioNRTJFgsUCK09D7dAewLjtT3WndKgyBZ7uA8lq1tVO9bvzfnCBUZHf4bprWbP6BO8T2H7TbOfQm297Gr8yvfa9y35GLVDD8cjVFenU-g_ezC8zT3wMT1zg</recordid><startdate>19990105</startdate><enddate>19990105</enddate><creator>Berry, J. 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Technologies</topic><topic>Microbiology</topic><topic>microcarrier</topic><topic>Microspheres</topic><topic>Orthoreovirus - physiology</topic><topic>reovirus</topic><topic>Reovirus 1</topic><topic>Reovirus 2</topic><topic>Vero</topic><topic>Vero Cells</topic><topic>Viral Plaque Assay</topic><topic>Viral Vaccines - isolation &amp; purification</topic><topic>Virus Cultivation - methods</topic><topic>Virus Replication</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Berry, J. M.</creatorcontrib><creatorcontrib>Barnabé, N.</creatorcontrib><creatorcontrib>Coombs, K. 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M.</au><au>Butler, M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Production of reovirus type-1 and type-3 from Vero cells grown on solid and macroporous microcarriers</atitle><jtitle>Biotechnology and bioengineering</jtitle><addtitle>Biotechnol. Bioeng</addtitle><date>1999-01-05</date><risdate>1999</risdate><volume>62</volume><issue>1</issue><spage>12</spage><epage>19</epage><pages>12-19</pages><issn>0006-3592</issn><eissn>1097-0290</eissn><coden>BIBIAU</coden><abstract>Two strains of reovirus were propagated in Vero cells grown in stationary or microcarriers cultures. Vero cells grown as monolayers on T‐flasks or in spinner cultures of Cytodex‐1 or Cultispher‐G microcarriers could be infected with reovirus serotype 1, strain Lang (T1L), and serotype 3, strain Dearing (T3D). A regime of intermittent low speed stirring at reduced culture volume was critical to ensure viral infection of cells in microcarrier cultures. The virus titre increased by 3 to 4 orders of magnitude over a culture period of 150 h. Titres of the T3D reovirus strain were higher (43%) compared to those of the T1L strain in all cultures. Titres were significantly higher in T‐flask and Cytodex‐1 microcarrier cultures compared to Cultispher‐G cultures with respect to either reovirus type. The viral productivity in the microcarrier cultures was dependent upon the multiplicity of infection (MOI) and the cell/bead ratio at the point of infection. A combination of high MOI (5 pfu/cell) and high cell/bead loading (&gt;400 for Cytodex‐1 and &gt;1,000 for Cultispher‐G) resulted in a low virus productivity per cell. However, at low MOI (0.5 pfu/cell) the virus productivity per cell was significantly higher at high cell/bead loading in cultures of either microcarrier type. The maximum virus titre (8.5 × 109 pfu/mL) was obtained in Cytodex‐1 cultures with a low MOI (0.5 pfu/cell) and a cell/bead loading of 1,000. The virus productivity per cell in these cultures was 4,000 pfu/cell. The lower viral yield in the Cultispher‐G microcarrier cultures is attributed to a decreased accessibility of the entrapped cells to viral infection. The high viral productivity from the Vero cells in Cytodex‐1 cultures suggests that this is a suitable system for the development of a vaccine production system for the Reoviridae viruses. © 1999 John Wiley &amp; Sons, Inc. Biotechnol Bioeng 62: 12–19, 1999.</abstract><cop>New York</cop><pub>John Wiley &amp; Sons, Inc</pub><pmid>10099508</pmid><doi>10.1002/(SICI)1097-0290(19990105)62:1&lt;12::AID-BIT2&gt;3.0.CO;2-G</doi><tpages>8</tpages></addata></record>
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subjects Animal cells
Animals
Biological and medical sciences
Biotechnology
Cercopithecus aethiops
Culture Media
Establishment of new cell lines, improvement of cultural methods, mass cultures
Eukaryotic cell cultures
Evaluation Studies as Topic
Fundamental and applied biological sciences. Psychology
Growth kinetics
macroporous
Mammalian orthoreovirus 3 - physiology
Methods. Procedures. Technologies
Microbiology
microcarrier
Microspheres
Orthoreovirus - physiology
reovirus
Reovirus 1
Reovirus 2
Vero
Vero Cells
Viral Plaque Assay
Viral Vaccines - isolation & purification
Virus Cultivation - methods
Virus Replication
Viruses
title Production of reovirus type-1 and type-3 from Vero cells grown on solid and macroporous microcarriers
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