Application of Multivariate Analysis to Optimize Function of Cultured Hepatocytes

Authors

  • Christina Chan,

    Corresponding author
    1. Department of Chemical Engineering and Material Science, Michigan State University, East Lansing, Michigan, 48824
    • Department of Chemical Engineering and Material Science, Michigan State University, East Lansing, Michigan, 48824. Tel: (517) 432–4530. Fax: (517) 432–1105
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  • Daehee Hwang,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge Massachusetts 02139
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  • Gregory N. Stephanopoulos,

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge Massachusetts 02139
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  • Martin L. Yarmush,

    1. Center for Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School and The Shriners Hospitals for Children, Boston, Massachusetts 02114
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  • George Stephanopoulos

    1. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge Massachusetts 02139
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Abstract

Understanding the metabolic and regulatory pathways of hepatocytes is important for biotechnological applications involving liver cells, including the development of bioartificial liver (BAL) devices. To characterize intermediary metabolism in the hepatocytes, metabolic flux analysis (MFA) was applied to elucidate the changes in intracellular pathway fluxes of primary rat hepatocytes exposed to human plasma and to provide a comprehensive snapshot of the hepatic metabolic profile. In the current study, the combination of preconditioning and plasma supplementation produced distinct metabolic states. Combining the metabolic flux distribution obtained by MFA with methodologies such as Fisher discriminant analysis (FDA) and partial least squares or projection to latent structures (PLS) provided insights into the underlying structure and causal relationship within the data. With the aid of these analyses, patterns in the cellular response of the hepatocytes that contributed to the separation of the different hepatic states were identified. Of particular interest was the recognition of distal pathways that strongly correlated with a particular hepatic function. The hepatic functions investigated were intracellular triglyceride accumulation and urea production. This study illustrates a framework for optimizing hepatic function and a possibility of identifying potential targets for improving hepatic functions.

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