High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor
Hepatitis C virus (HCV) infections pose a major public health burden due to high chronicity rates and associated morbidity and mortality. A vaccine protecting against chronic infection is not available but would be important for global control of HCV infections. In this study, cell culture-based HCV...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-02-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/10/2/249 |
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| author | Anna Offersgaard Carlos Rene Duarte Hernandez Anne Finne Pihl Nandini Prabhakar Venkatesan Henrik Krarup Xiangliang Lin Udo Reichl Jens Bukh Yvonne Genzel Judith Margarete Gottwein |
| author_facet | Anna Offersgaard Carlos Rene Duarte Hernandez Anne Finne Pihl Nandini Prabhakar Venkatesan Henrik Krarup Xiangliang Lin Udo Reichl Jens Bukh Yvonne Genzel Judith Margarete Gottwein |
| author_sort | Anna Offersgaard |
| collection | DOAJ |
| description | Hepatitis C virus (HCV) infections pose a major public health burden due to high chronicity rates and associated morbidity and mortality. A vaccine protecting against chronic infection is not available but would be important for global control of HCV infections. In this study, cell culture-based HCV production was established in a packed-bed bioreactor (CelCradle™) aiming to further the development of an inactivated whole virus vaccine and to facilitate virological and immunological studies requiring large quantities of virus particles. HCV was produced in human hepatoma-derived Huh7.5 cells maintained in serum-free medium on days of virus harvesting. Highest virus yields were obtained when the culture was maintained with two medium exchanges per day. However, increasing the total number of cells in the culture vessel negatively impacted infectivity titers. Peak infectivity titers of up to 7.2 log<sub>10</sub> focus forming units (FFU)/mL, accumulated virus yields of up to 5.9 × 10<sup>10</sup> FFU, and a cell specific virus yield of up to 41 FFU/cell were obtained from one CelCradle™. CelCradle™-derived and T flask-derived virus had similar characteristics regarding neutralization sensitivity and buoyant density. This packed-bed tide-motion system is available with larger vessels and may thus be a promising platform for large-scale HCV production. |
| first_indexed | 2024-03-09T20:54:39Z |
| format | Article |
| id | doaj.art-936ce43852514f91a6740978172983f3 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-09T20:54:39Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-936ce43852514f91a6740978172983f32023-11-23T22:25:58ZengMDPI AGVaccines2076-393X2022-02-0110224910.3390/vaccines10020249High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density BioreactorAnna Offersgaard0Carlos Rene Duarte Hernandez1Anne Finne Pihl2Nandini Prabhakar Venkatesan3Henrik Krarup4Xiangliang Lin5Udo Reichl6Jens Bukh7Yvonne Genzel8Judith Margarete Gottwein9Copenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Copenhagen University Hospital–Hvidovre, 2650 Hvidovre, DenmarkCopenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Copenhagen University Hospital–Hvidovre, 2650 Hvidovre, DenmarkCopenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Copenhagen University Hospital–Hvidovre, 2650 Hvidovre, DenmarkEsco Aster Pte Ltd., Singapore 486 777, SingaporeDepartment of Molecular Diagnostics, Aalborg University Hospital, 9000 Aalborg, DenmarkEsco Aster Pte Ltd., Singapore 486 777, SingaporeBioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, GermanyCopenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Copenhagen University Hospital–Hvidovre, 2650 Hvidovre, DenmarkBioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, GermanyCopenhagen Hepatitis C Program (CO-HEP), Department of Infectious Diseases, Copenhagen University Hospital–Hvidovre, 2650 Hvidovre, DenmarkHepatitis C virus (HCV) infections pose a major public health burden due to high chronicity rates and associated morbidity and mortality. A vaccine protecting against chronic infection is not available but would be important for global control of HCV infections. In this study, cell culture-based HCV production was established in a packed-bed bioreactor (CelCradle™) aiming to further the development of an inactivated whole virus vaccine and to facilitate virological and immunological studies requiring large quantities of virus particles. HCV was produced in human hepatoma-derived Huh7.5 cells maintained in serum-free medium on days of virus harvesting. Highest virus yields were obtained when the culture was maintained with two medium exchanges per day. However, increasing the total number of cells in the culture vessel negatively impacted infectivity titers. Peak infectivity titers of up to 7.2 log<sub>10</sub> focus forming units (FFU)/mL, accumulated virus yields of up to 5.9 × 10<sup>10</sup> FFU, and a cell specific virus yield of up to 41 FFU/cell were obtained from one CelCradle™. CelCradle™-derived and T flask-derived virus had similar characteristics regarding neutralization sensitivity and buoyant density. This packed-bed tide-motion system is available with larger vessels and may thus be a promising platform for large-scale HCV production.https://www.mdpi.com/2076-393X/10/2/249packed-bed bioreactorCelCradle™high cell density cell cultureHuh7.5 cellswhole virus vaccineinactivated virus vaccine |
| spellingShingle | Anna Offersgaard Carlos Rene Duarte Hernandez Anne Finne Pihl Nandini Prabhakar Venkatesan Henrik Krarup Xiangliang Lin Udo Reichl Jens Bukh Yvonne Genzel Judith Margarete Gottwein High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor Vaccines packed-bed bioreactor CelCradle™ high cell density cell culture Huh7.5 cells whole virus vaccine inactivated virus vaccine |
| title | High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor |
| title_full | High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor |
| title_fullStr | High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor |
| title_full_unstemmed | High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor |
| title_short | High-Titer Hepatitis C Virus Production in a Scalable Single-Use High Cell Density Bioreactor |
| title_sort | high titer hepatitis c virus production in a scalable single use high cell density bioreactor |
| topic | packed-bed bioreactor CelCradle™ high cell density cell culture Huh7.5 cells whole virus vaccine inactivated virus vaccine |
| url | https://www.mdpi.com/2076-393X/10/2/249 |
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