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Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy

BACKGROUND: Adoptive T-cell therapy is a promising treatment strategy for cancer immunotherapy. The ability of immunotherapy based on the adoptive cell transfer of genetically modified T cells to generate powerful clinical responses has been highlighted by recent clinical success. Techniques which a...

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Autores principales: Rahimmanesh, Ilnaz, Tavangar, Mehrsa, Zahedi, Seyedeh Noushin, Azizi, Yadollah, Khanahmad Shahreza, Hossein
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Wolters Kluwer - Medknow 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9744083/
https://www.ncbi.nlm.nih.gov/pubmed/36518860
http://dx.doi.org/10.4103/abr.abr_349_21
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author Rahimmanesh, Ilnaz
Tavangar, Mehrsa
Zahedi, Seyedeh Noushin
Azizi, Yadollah
Khanahmad Shahreza, Hossein
author_facet Rahimmanesh, Ilnaz
Tavangar, Mehrsa
Zahedi, Seyedeh Noushin
Azizi, Yadollah
Khanahmad Shahreza, Hossein
author_sort Rahimmanesh, Ilnaz
collection PubMed
description BACKGROUND: Adoptive T-cell therapy is a promising treatment strategy for cancer immunotherapy. The ability of immunotherapy based on the adoptive cell transfer of genetically modified T cells to generate powerful clinical responses has been highlighted by recent clinical success. Techniques which are used to expand large numbers of T cells from different sources are critical in adoptive cell therapy. In this study, we evaluated the expansion, proliferation, activation of T lymphocytes, in the presence of various concentrations of interleukin-2, phytohemagglutinin (PHA), and insulin. MATERIALS AND METHODS: The effect of different supplemented culture media on T cell expansion was evaluated using MTT assay. The expression level of the Ki-67 proliferation marker was evaluated by real-time polymerase chain reaction. In addition, flow cytometry analysis was performed to access T cell subpopulations. RESULTS: Our results showed that supplemented culture media with an optimized concentration of PHA and interleukin-2 increased total fold expansion of T cells up to 500-fold with approximately 90% cell viability over 7 days. The quantitative assessment of Ki-67 in expanded T cells showed a significant elevation of this proliferation marker. Flow cytometry was also used to assess the proportion of CD4+ and CD8+ cells, and the main expanded population was CD3+ CD8+ cells. CONCLUSIONS: Based on these findings, we introduced a low-cost and rapid method to support the efficient expansion of T cells for adoptive cell therapy and other in vivo experiments.
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spelling pubmed-97440832022-12-13 Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy Rahimmanesh, Ilnaz Tavangar, Mehrsa Zahedi, Seyedeh Noushin Azizi, Yadollah Khanahmad Shahreza, Hossein Adv Biomed Res Original Article BACKGROUND: Adoptive T-cell therapy is a promising treatment strategy for cancer immunotherapy. The ability of immunotherapy based on the adoptive cell transfer of genetically modified T cells to generate powerful clinical responses has been highlighted by recent clinical success. Techniques which are used to expand large numbers of T cells from different sources are critical in adoptive cell therapy. In this study, we evaluated the expansion, proliferation, activation of T lymphocytes, in the presence of various concentrations of interleukin-2, phytohemagglutinin (PHA), and insulin. MATERIALS AND METHODS: The effect of different supplemented culture media on T cell expansion was evaluated using MTT assay. The expression level of the Ki-67 proliferation marker was evaluated by real-time polymerase chain reaction. In addition, flow cytometry analysis was performed to access T cell subpopulations. RESULTS: Our results showed that supplemented culture media with an optimized concentration of PHA and interleukin-2 increased total fold expansion of T cells up to 500-fold with approximately 90% cell viability over 7 days. The quantitative assessment of Ki-67 in expanded T cells showed a significant elevation of this proliferation marker. Flow cytometry was also used to assess the proportion of CD4+ and CD8+ cells, and the main expanded population was CD3+ CD8+ cells. CONCLUSIONS: Based on these findings, we introduced a low-cost and rapid method to support the efficient expansion of T cells for adoptive cell therapy and other in vivo experiments. Wolters Kluwer - Medknow 2022-10-31 /pmc/articles/PMC9744083/ /pubmed/36518860 http://dx.doi.org/10.4103/abr.abr_349_21 Text en Copyright: © 2022 Advanced Biomedical Research https://creativecommons.org/licenses/by-nc-sa/4.0/This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.
spellingShingle Original Article
Rahimmanesh, Ilnaz
Tavangar, Mehrsa
Zahedi, Seyedeh Noushin
Azizi, Yadollah
Khanahmad Shahreza, Hossein
Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title_full Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title_fullStr Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title_full_unstemmed Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title_short Optimization of Culture Media for Ex vivo T-Cell Expansion for Adoptive T-Cell Therapy
title_sort optimization of culture media for ex vivo t-cell expansion for adoptive t-cell therapy
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9744083/
https://www.ncbi.nlm.nih.gov/pubmed/36518860
http://dx.doi.org/10.4103/abr.abr_349_21
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