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Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes

Cell detachment techniques using animal-derived enzymes are necessary for the production of biopharmaceuticals that are made with the help of adherent cell cultures, although the majority of protein therapeutics (>USD 100 billion of income per year) are made under suspension cultures that do not...

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Autores principales: Yu, Shouzhi, Huang, Yunchao, Wu, Chongyang, Fu, Weibin, Liang, Hongyang, Chen, Chen, Cheng, Yue, Guo, Yancen, Zhang, Ying, Wang, Hui, Yang, Xiaoming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9656133/
https://www.ncbi.nlm.nih.gov/pubmed/36359792
http://dx.doi.org/10.3390/cells11213396
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author Yu, Shouzhi
Huang, Yunchao
Wu, Chongyang
Fu, Weibin
Liang, Hongyang
Chen, Chen
Cheng, Yue
Guo, Yancen
Zhang, Ying
Wang, Hui
Yang, Xiaoming
author_facet Yu, Shouzhi
Huang, Yunchao
Wu, Chongyang
Fu, Weibin
Liang, Hongyang
Chen, Chen
Cheng, Yue
Guo, Yancen
Zhang, Ying
Wang, Hui
Yang, Xiaoming
author_sort Yu, Shouzhi
collection PubMed
description Cell detachment techniques using animal-derived enzymes are necessary for the production of biopharmaceuticals that are made with the help of adherent cell cultures, although the majority of protein therapeutics (>USD 100 billion of income per year) are made under suspension cultures that do not require animal-derived proteins for manufacture. In this study, we establish the optimal Vero cell detachment process, and analyze physiological changes during cell detachment at the cellular and molecular levels. Using flow cytometry, we find that animal-based enzymes are more likely to induce apoptosis than animal-origin-free enzymes. We analyze the levels of RNAs, proteins, and metabolites in cells treated with two detachment strategies, and identify 1237 differentially expressed genes, 2883 differential proteins, and 210 differential metabolites. Transcriptomic analysis shows that animal-origin-free enzymes have a less significant effect on gene expression levels. Combined with proteomic analysis, animal-based enzymes affect the oxidative phosphorylation process and reduce the mRNA and protein levels of Cytochrome C Oxidase Assembly Protein 17 (COX17), which is a Cytochrome C Oxidase Copper Chaperone involved in the mitochondrial respiratory chain. Metabolomics analysis indicates that the levels of spermine and spermidine, which are involved in the glutathione metabolism pathway and apoptosis inhibition, are significantly reduced. Therefore, COX17, spermine, and spermidine may be biomarkers for evaluating the cell subculture process. In conclusion, we have deeply characterized the cell subculture process through multi-omics, which may provide important guidance for research and process evaluation to optimize cell detachment processes.
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spelling pubmed-96561332022-11-15 Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes Yu, Shouzhi Huang, Yunchao Wu, Chongyang Fu, Weibin Liang, Hongyang Chen, Chen Cheng, Yue Guo, Yancen Zhang, Ying Wang, Hui Yang, Xiaoming Cells Article Cell detachment techniques using animal-derived enzymes are necessary for the production of biopharmaceuticals that are made with the help of adherent cell cultures, although the majority of protein therapeutics (>USD 100 billion of income per year) are made under suspension cultures that do not require animal-derived proteins for manufacture. In this study, we establish the optimal Vero cell detachment process, and analyze physiological changes during cell detachment at the cellular and molecular levels. Using flow cytometry, we find that animal-based enzymes are more likely to induce apoptosis than animal-origin-free enzymes. We analyze the levels of RNAs, proteins, and metabolites in cells treated with two detachment strategies, and identify 1237 differentially expressed genes, 2883 differential proteins, and 210 differential metabolites. Transcriptomic analysis shows that animal-origin-free enzymes have a less significant effect on gene expression levels. Combined with proteomic analysis, animal-based enzymes affect the oxidative phosphorylation process and reduce the mRNA and protein levels of Cytochrome C Oxidase Assembly Protein 17 (COX17), which is a Cytochrome C Oxidase Copper Chaperone involved in the mitochondrial respiratory chain. Metabolomics analysis indicates that the levels of spermine and spermidine, which are involved in the glutathione metabolism pathway and apoptosis inhibition, are significantly reduced. Therefore, COX17, spermine, and spermidine may be biomarkers for evaluating the cell subculture process. In conclusion, we have deeply characterized the cell subculture process through multi-omics, which may provide important guidance for research and process evaluation to optimize cell detachment processes. MDPI 2022-10-27 /pmc/articles/PMC9656133/ /pubmed/36359792 http://dx.doi.org/10.3390/cells11213396 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Yu, Shouzhi
Huang, Yunchao
Wu, Chongyang
Fu, Weibin
Liang, Hongyang
Chen, Chen
Cheng, Yue
Guo, Yancen
Zhang, Ying
Wang, Hui
Yang, Xiaoming
Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title_full Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title_fullStr Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title_full_unstemmed Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title_short Integrated Multi-Omic Characterization of the Detachment Process of Adherent Vero Cells with Animal-Based and Animal-Origin-Free Enzymes
title_sort integrated multi-omic characterization of the detachment process of adherent vero cells with animal-based and animal-origin-free enzymes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9656133/
https://www.ncbi.nlm.nih.gov/pubmed/36359792
http://dx.doi.org/10.3390/cells11213396
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