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Untargeted LC-MS Metabolomics for the Analysis of Micro-scaled Extracellular Metabolites from Hepatocytes

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Abstract

Metabolome analysis in micro physiological models is a challenge due to the low volume of the cell culture medium (CCM). Here, we report a LC-MS-based untargeted metabolomics protocol for the detection of hepatocyte extracellular metabolites from micro-scale samples of CCM. Using a single LC-MS method we have detected 57 metabolites of which 27 showed >2-fold shifts after 72-hour incubation. We demonstrate that micro-scale CCM samples can be used for modelling micro-physiological temporal dynamics in metabolite intensities.

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Acknowledgements

This work was supported by JSPS KAKENHI Grant Number 17H02083, 20K20168 and 19K17662. Kyoto University GAP fund program (207010). We acknowledge the support from the Gunma University Initiative for Advanced Research (GIAR). I. M. was supported by Japan Society for the Promotion of Science (JSPS) postdoctoral fellowship (P17774). Authors thank Dr. Satoshi Imamura for assisting in HepG2 cells culture.

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Authors and Affiliations

  1. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-Ushinomiya-cho, Sakyo, Kyoto, 606-8501, Japan

    Rodi Abdalkader & Ken-ichiro Kamei

  2. Gunma University Initiative for Advanced Research (GIAR), Gunma University, Gunma, 371-8511, Japan

    Romanas Chaleckis, Isabel Meister, Pei Zhang & Craig E. Wheelock

  3. Division of Physiological Chemistry II, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden

    Romanas Chaleckis, Isabel Meister, Pei Zhang & Craig E. Wheelock

  4. Wuya College of Innovation, Shenyang Pharmaceutical University, Liaoning, 110016, China

    Ken-ichiro Kamei

Authors
  1. Rodi Abdalkader
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  2. Romanas Chaleckis
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  3. Isabel Meister
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  4. Pei Zhang
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  5. Craig E. Wheelock
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  6. Ken-ichiro Kamei
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Correspondence to Rodi Abdalkader.

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Abdalkader, R., Chaleckis, R., Meister, I. et al. Untargeted LC-MS Metabolomics for the Analysis of Micro-scaled Extracellular Metabolites from Hepatocytes. ANAL. SCI. 37, 1049–1052 (2021). https://doi.org/10.2116/analsci.20N032

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  • DOI: https://doi.org/10.2116/analsci.20N032

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