Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media

An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system. In this study, global metabolite (metabolomics) analysis approach was used to try a...

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Main Authors:	Mohamad Saberi, Salfarina Ezrina, Hashim, Yumi Zuhanis Has-Yun, Mel, Maizirwan, Amid, Azura, Ahmad-Raus, Raha, Packeer Mohamed, Vasila
Format:	Article
Language:	English English
Published:	Springer 2013
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/1/metabolomics.pdf http://irep.iium.edu.my/27589/4/Yumi_Metabolics_Profiling.pdf

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spelling	iium-275892013-12-17T02:00:18Z http://irep.iium.edu.my/27589/ Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media Mohamad Saberi, Salfarina Ezrina Hashim, Yumi Zuhanis Has-Yun Mel, Maizirwan Amid, Azura Ahmad-Raus, Raha Packeer Mohamed, Vasila Q Science (General) An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system. In this study, global metabolite (metabolomics) analysis approach was used to try and understand the relationships between types of media used, culture growth behavior and productivity. CHO-KI cells producing IGF-1 were obtained from ATCC and grown in T-flask (37 °C, 5 % CO2) until 70–80 % confluent in RPMI 1640 and Ham’s F12, respectively. Samples were taken at 8-hourly intervals for routine cell counting, biochemical responses, insulin like growth factor—1 (IGF-1) protein concentration and global metabolite analysis (gas chromatography mass spectrometry, GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCA-P + Version 12 for chemometric evaluation using principal component analysis. The results showed that while routine analysis gave only subtle differences between the media, global metabolite analysis was able to clearly separate the culture based on growth media with growth phases as confounding factor. Different types of media also appeared to affect IGF-1 production. Asparagine was found to be indicative of healthiness of cells and production of high IGF-1. Meanwhile identification of ornithine and lysine in death phase was found to be associated with apoptosis and oversupplied nutrient respectively. Using the biomarkers revealed in the study, several bioprocessing strategies including medium improvement and in-time downstream processing can be potentially implemented to achieve efficient CHO culture system. Springer 2013-08 Article PeerReviewed application/pdf en http://irep.iium.edu.my/27589/1/metabolomics.pdf application/pdf en http://irep.iium.edu.my/27589/4/Yumi_Metabolics_Profiling.pdf Mohamad Saberi, Salfarina Ezrina and Hashim, Yumi Zuhanis Has-Yun and Mel, Maizirwan and Amid, Azura and Ahmad-Raus, Raha and Packeer Mohamed, Vasila (2013) Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media. Cytotechnology, 65 (4). pp. 577-586. ISSN 1573-0778 (O), 0920-9069 (P) http://link.springer.com/content/pdf/10.1007%2Fs10616-012-9508-4 10.1007/s10616-012-9508-4
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	Q Science (General)
spellingShingle	Q Science (General) Mohamad Saberi, Salfarina Ezrina Hashim, Yumi Zuhanis Has-Yun Mel, Maizirwan Amid, Azura Ahmad-Raus, Raha Packeer Mohamed, Vasila Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
description	An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system. In this study, global metabolite (metabolomics) analysis approach was used to try and understand the relationships between types of media used, culture growth behavior and productivity. CHO-KI cells producing IGF-1 were obtained from ATCC and grown in T-flask (37 °C, 5 % CO2) until 70–80 % confluent in RPMI 1640 and Ham’s F12, respectively. Samples were taken at 8-hourly intervals for routine cell counting, biochemical responses, insulin like growth factor—1 (IGF-1) protein concentration and global metabolite analysis (gas chromatography mass spectrometry, GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCA-P + Version 12 for chemometric evaluation using principal component analysis. The results showed that while routine analysis gave only subtle differences between the media, global metabolite analysis was able to clearly separate the culture based on growth media with growth phases as confounding factor. Different types of media also appeared to affect IGF-1 production. Asparagine was found to be indicative of healthiness of cells and production of high IGF-1. Meanwhile identification of ornithine and lysine in death phase was found to be associated with apoptosis and oversupplied nutrient respectively. Using the biomarkers revealed in the study, several bioprocessing strategies including medium improvement and in-time downstream processing can be potentially implemented to achieve efficient CHO culture system.
format	Article
author	Mohamad Saberi, Salfarina Ezrina Hashim, Yumi Zuhanis Has-Yun Mel, Maizirwan Amid, Azura Ahmad-Raus, Raha Packeer Mohamed, Vasila
author_facet	Mohamad Saberi, Salfarina Ezrina Hashim, Yumi Zuhanis Has-Yun Mel, Maizirwan Amid, Azura Ahmad-Raus, Raha Packeer Mohamed, Vasila
author_sort	Mohamad Saberi, Salfarina Ezrina
title	Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
title_short	Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
title_full	Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
title_fullStr	Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
title_full_unstemmed	Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media
title_sort	metabolomics profiling of extracellular metabolites in cho-k1 cells cultured in different types of growth media
publisher	Springer
publishDate	2013
url	http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/ http://irep.iium.edu.my/27589/1/metabolomics.pdf http://irep.iium.edu.my/27589/4/Yumi_Metabolics_Profiling.pdf
first_indexed	2023-09-18T20:40:57Z
last_indexed	2023-09-18T20:40:57Z
_version_	1777409361171185664

Metabolomics profiling of extracellular metabolites in CHO-K1 cells cultured in different types of growth media

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