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Medicine
Aislamiento de hepatocitos regeneradores después de hepatectomía parcial en ratones
Aislamiento de hepatocitos regeneradores después de hepatectomía parcial en ratones
JoVE Journal
Medicine
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JoVE Journal Medicine
Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

Aislamiento de hepatocitos regeneradores después de hepatectomía parcial en ratones

Full Text
5,762 Views
10:04 min
December 2, 2022

DOI: 10.3791/64493-v

Eva Breuer1, Bostjan Humar1

1Swiss HPB Laboratory, Department of Visceral & Transplant Surgery,University Hospital Zurich (USZ)

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Please note that some of the translations on this page are AI generated. Click here for the English version.

Overview

This article presents an optimized protocol for isolating hepatocytes in mice following partial hepatectomy. The method effectively retains lipid-laden hepatocytes, which are typically lost during density-gradient centrifugation, allowing for the study of regenerating hepatocyte populations.

Key Study Components

Area of Science

  • Cell biology
  • Liver regeneration
  • Hepatocyte isolation

Background

  • Lipid-laden hepatocytes are crucial for liver regeneration.
  • Density-gradient centrifugation often results in the loss of these cells.
  • Optimizing isolation methods can enhance research on liver regeneration.
  • This study focuses on improving hepatocyte recovery post-surgery.

Purpose of Study

  • To develop a protocol that retains steatotic hepatocytes.
  • To provide a reliable method for studying regenerating hepatocytes.
  • To eliminate the need for density-gradient centrifugation in hepatocyte isolation.

Methods Used

  • Preparation of perfusion equipment using a 26 gauge IUI cannula.
  • Flushing and priming the tubing with warm perfusion buffer.
  • Maintaining a pump speed of three milliliters per minute.
  • Isolating hepatocytes post-partial hepatectomy in mice.

Main Results

  • The protocol successfully retains lipid-laden hepatocytes.
  • Isolated hepatocytes represent a more accurate population for study.
  • Enhanced recovery of regenerating hepatocytes was observed.
  • The method shows promise for future liver regeneration research.

Conclusions

  • The optimized isolation protocol is effective for hepatocyte recovery.
  • Retaining steatotic hepatocytes is crucial for studying liver regeneration.
  • This method can facilitate further research in liver biology.

Frequently Asked Questions

What is the significance of lipid-laden hepatocytes?
Lipid-laden hepatocytes play a critical role in liver regeneration and metabolic processes.
How does this protocol differ from traditional methods?
This protocol eliminates the need for density-gradient centrifugation, improving the retention of specific cell types.
What are the potential applications of this research?
The findings can be applied to studies on liver diseases, regeneration, and metabolic disorders.
Can this method be used in other animal models?
While this study focuses on mice, adaptations may be possible for other species.
What are the next steps for this research?
Future studies may explore the functional characteristics of the isolated hepatocytes.
Is this protocol applicable for clinical research?
The protocol may provide insights that could be relevant for clinical applications in liver health.

Los hepatocitos cargados de lípidos son inherentes a la regeneración hepática, pero generalmente se pierden con la centrifugación por gradiente de densidad. Aquí, presentamos un protocolo de aislamiento celular optimizado que retiene hepatocitos esteatósicos, produciendo poblaciones representativas de hepatocitos regeneradores después de la hepatectomía parcial en ratones.

El objetivo general de este protocolo es presentar un método optimizado para el aislamiento de hepatocitos en ratones después de una hepatectomía parcial sin necesidad de centrifugación por gradiente de densidad. Por lo general, vemos un gran porcentaje de hepatocitos cargados de lípidos durante la regeneración temprana. Los hepatocitos esteatósicos que generalmente se pierden en la centrifugación del gradiente de densidad debido a la baja densidad se conservan con este protocolo.

Para preparar el equipo de perfusión, conecte una cánula IUI de calibre 26 al extremo de salida del tubo mediante un conector de bloqueo Luer. Luego enjuague el tubo e inserte el extremo de entrada del tubo en el tubo tampón de perfusión precalentado en el baño de agua. Prepárelo con tampón de perfusión caliente a una velocidad de bombeo de tres mililitros por minuto.

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