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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/27589/1/metabolomics.pdf http://irep.iium.edu.my/27589/4/Yumi_Metabolics_Profiling.pdf http://irep.iium.edu.my/27589/ http://link.springer.com/content/pdf/10.1007%2Fs10616-012-9508-4
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spelling	my.iium.irep.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 REM 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
institution	Universiti Islam Antarabangsa Malaysia
building	IIUM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	International Islamic University Malaysia
content_source	IIUM Repository (IREP)
url_provider	http://irep.iium.edu.my/
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/27589/1/metabolomics.pdf http://irep.iium.edu.my/27589/4/Yumi_Metabolics_Profiling.pdf http://irep.iium.edu.my/27589/ http://link.springer.com/content/pdf/10.1007%2Fs10616-012-9508-4
_version_	1643609361524719616
score	13.211869

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

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