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Related Experiment Videos

A new method for determining cellular ATP content in hepatocytes

M Ishikawa1, S Yogita, S Takai

  • 11st Department of Surgery, School of Medicine, University of Tokushima, Japan.

The Tokushima Journal of Experimental Medicine
|December 1, 1993
PubMed
Summary

A new adenosine triphosphate (ATP) assay for hepatocytes is simple, rapid, and sensitive. This method accurately measures ATP levels in viable cells and shows changes in liver regeneration and after transcatheter portal embolization.

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Area of Science:

  • Biochemistry
  • Hepatology
  • Cell Biology

Background:

  • Accurate measurement of cellular adenosine triphosphate (ATP) is crucial for assessing hepatocyte viability and function.
  • Existing methods for ATP determination can be complex or lack sensitivity.
  • Hepatocyte ATP levels reflect cellular metabolic status and response to physiological or pathological changes.

Purpose of the Study:

  • To develop and validate a novel, highly sensitive ATP assay for isolated hepatocytes.
  • To correlate hepatocyte ATP content with viable cell number.
  • To apply the assay to study ATP changes during liver regeneration and after transcatheter portal embolization (TPE).

Main Methods:

  • Hepatocytes were isolated using collagenase perfusion.
  • ATP was quantified using the luciferin-luciferase assay.

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  • Cell viability was assessed using the trypan blue exclusion test.
  • ATP levels were measured in rat hepatocytes post-partial hepatectomy and in dog hepatocytes post-TPE.

    • Main Results:

    • The developed ATP assay demonstrated high sensitivity and a strong correlation with viable hepatocyte numbers.
    • Dead hepatocytes showed minimal ATP content (<1% of viable cells).
    • Rat hepatocytes showed a significant increase in ATP content 24 hours after partial hepatectomy.
    • In dogs, TPE led to decreased ATP in embolized lobes and increased ATP in compensatory hypertrophied non-embolized lobes.

    Conclusions:

    • The luciferin-luciferase based ATP assay is a reliable and sensitive method for quantifying ATP in hepatocytes.
    • This assay is valuable for assessing hepatocyte viability and metabolic changes in response to liver injury and regeneration.
    • The method's adaptability may extend to the analysis of other adenosine phosphates.