Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells

Detalhes bibliográficos
Autor(a) principal: Rodrigues, E.
Data de Publicação: 2011
Outros Autores: Fernandes, Pedro, Costa, A. R., Henriques, Mariana, Azeredo, Joana, Oliveira, Rosário
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/1822/25772
Resumo: Large-scale biopharmaceutical production commonly relies on suspension cell cultures that provide higher yields than adherent cultures. However, most mammalian cells grow adherently and therefore need to be adapted to suspended growth, which is not always simple or feasible. Microcarrier culture introduces new possibilities and makes achievable the practical high yield culture of anchorage-dependent cells in suspension systems. The aim of this study was to evaluate and optimize the use of microcarrier culture for the growth and antibody production of CHO-K1 cells. For this, the macroporous Cultispher microcarriers were used, and the initial cell adhesion to the microcarriers (occurring in the first 5-6 hours) and further cell proliferation were assessed. Cultures of antibody-producing CHO-K1 cells were performed in 50 ml vented conical tubes, and different conditions were tested: initial cell concentration (2x105 cells/ml and 4x105 cells/ml), microcarrier concentration (1 g/L and 2 g/L), type of rocking during the first 6 hours of adhesion (pulse or continuous) and rocking after initial adhesion (no rocking and 60 rpm). Cell concentration and viability in the microcarriers were assessed periodically (hourly for the adhesion phase, and daily after that). It was observed that an increase in the initial cell concentration does not enhance initial adhesion, possibly due to saturation of the microcarrier surface. For its turn, increasing microcarrier concentration, without further increasing initial cell concentration does not improve cell densities achieved in the culture. Concerning rocking, the most favorable type for the adhesion phase was pulse rocking and, after this, a continuous rocking provided an improved cell proliferation. In conclusion, microcarrier cultures proved to be a viable alternative to suspended cultures for the growth and antibody production of CHO-K1 cells.
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spelling Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cellsLarge-scale biopharmaceutical production commonly relies on suspension cell cultures that provide higher yields than adherent cultures. However, most mammalian cells grow adherently and therefore need to be adapted to suspended growth, which is not always simple or feasible. Microcarrier culture introduces new possibilities and makes achievable the practical high yield culture of anchorage-dependent cells in suspension systems. The aim of this study was to evaluate and optimize the use of microcarrier culture for the growth and antibody production of CHO-K1 cells. For this, the macroporous Cultispher microcarriers were used, and the initial cell adhesion to the microcarriers (occurring in the first 5-6 hours) and further cell proliferation were assessed. Cultures of antibody-producing CHO-K1 cells were performed in 50 ml vented conical tubes, and different conditions were tested: initial cell concentration (2x105 cells/ml and 4x105 cells/ml), microcarrier concentration (1 g/L and 2 g/L), type of rocking during the first 6 hours of adhesion (pulse or continuous) and rocking after initial adhesion (no rocking and 60 rpm). Cell concentration and viability in the microcarriers were assessed periodically (hourly for the adhesion phase, and daily after that). It was observed that an increase in the initial cell concentration does not enhance initial adhesion, possibly due to saturation of the microcarrier surface. For its turn, increasing microcarrier concentration, without further increasing initial cell concentration does not improve cell densities achieved in the culture. Concerning rocking, the most favorable type for the adhesion phase was pulse rocking and, after this, a continuous rocking provided an improved cell proliferation. In conclusion, microcarrier cultures proved to be a viable alternative to suspended cultures for the growth and antibody production of CHO-K1 cells.BioMed Central (BMC)Universidade do MinhoRodrigues, E.Fernandes, PedroCosta, A. R.Henriques, MarianaAzeredo, JoanaOliveira, Rosário20112011-01-01T00:00:00Zconference objectinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/1822/25772enginfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-05-11T05:55:48Zoai:repositorium.sdum.uminho.pt:1822/25772Portal AgregadorONGhttps://www.rcaap.pt/oai/openairemluisa.alvim@gmail.comopendoar:71602024-05-11T05:55:48Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
title Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
spellingShingle Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
Rodrigues, E.
title_short Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
title_full Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
title_fullStr Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
title_full_unstemmed Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
title_sort Preliminary evaluation of microcarrier culture for growth and monoclonal antibody production of CHO-K1 cells
author Rodrigues, E.
author_facet Rodrigues, E.
Fernandes, Pedro
Costa, A. R.
Henriques, Mariana
Azeredo, Joana
Oliveira, Rosário
author_role author
author2 Fernandes, Pedro
Costa, A. R.
Henriques, Mariana
Azeredo, Joana
Oliveira, Rosário
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Rodrigues, E.
Fernandes, Pedro
Costa, A. R.
Henriques, Mariana
Azeredo, Joana
Oliveira, Rosário
description Large-scale biopharmaceutical production commonly relies on suspension cell cultures that provide higher yields than adherent cultures. However, most mammalian cells grow adherently and therefore need to be adapted to suspended growth, which is not always simple or feasible. Microcarrier culture introduces new possibilities and makes achievable the practical high yield culture of anchorage-dependent cells in suspension systems. The aim of this study was to evaluate and optimize the use of microcarrier culture for the growth and antibody production of CHO-K1 cells. For this, the macroporous Cultispher microcarriers were used, and the initial cell adhesion to the microcarriers (occurring in the first 5-6 hours) and further cell proliferation were assessed. Cultures of antibody-producing CHO-K1 cells were performed in 50 ml vented conical tubes, and different conditions were tested: initial cell concentration (2x105 cells/ml and 4x105 cells/ml), microcarrier concentration (1 g/L and 2 g/L), type of rocking during the first 6 hours of adhesion (pulse or continuous) and rocking after initial adhesion (no rocking and 60 rpm). Cell concentration and viability in the microcarriers were assessed periodically (hourly for the adhesion phase, and daily after that). It was observed that an increase in the initial cell concentration does not enhance initial adhesion, possibly due to saturation of the microcarrier surface. For its turn, increasing microcarrier concentration, without further increasing initial cell concentration does not improve cell densities achieved in the culture. Concerning rocking, the most favorable type for the adhesion phase was pulse rocking and, after this, a continuous rocking provided an improved cell proliferation. In conclusion, microcarrier cultures proved to be a viable alternative to suspended cultures for the growth and antibody production of CHO-K1 cells.
publishDate 2011
dc.date.none.fl_str_mv 2011
2011-01-01T00:00:00Z
dc.type.driver.fl_str_mv conference object
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status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/1822/25772
url http://hdl.handle.net/1822/25772
dc.language.iso.fl_str_mv eng
language eng
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dc.publisher.none.fl_str_mv BioMed Central (BMC)
publisher.none.fl_str_mv BioMed Central (BMC)
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv mluisa.alvim@gmail.com
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