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Medicine
Isolamento de Hepatócitos Regeneradores após Hepatectomia Parcial em Camundongos
Isolamento de Hepatócitos Regeneradores após Hepatectomia Parcial em Camundongos
JoVE Journal
Medicine
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JoVE Journal Medicine
Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

Isolamento de Hepatócitos Regeneradores após Hepatectomia Parcial em Camundongos

Full Text
5,774 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.

Hepatócitos carregados de lipídios são inerentes à regeneração hepática, mas geralmente são perdidos após a centrifugação do gradiente de densidade. Aqui, apresentamos um protocolo de isolamento celular otimizado que retém hepatócitos esteatóticos, produzindo populações representativas de hepatócitos regeneradores após hepatectomia parcial em camundongos.

O objetivo geral deste protocolo é apresentar um método otimizado para o isolamento de hepatócitos em camundongos após hepatectomia parcial sem a necessidade de centrifugação por gradiente de densidade. Geralmente vemos uma grande porcentagem de hepatócitos carregados de lipídios durante a regeneração precoce. Os hepatócitos esteatóticos geralmente perdidos após a centrifugação do gradiente de densidade devido à baixa densidade são mantidos com este protocolo.

Para preparar o equipamento de perfusão, conecte uma cânula IUI de calibre 26 à extremidade de saída da tubulação usando um conector de bloqueio Luer. Em seguida, lave a tubulação e insira a extremidade de entrada do tubo no tubo tampão de perfusão pré-aquecido no banho de água. Prime-o com tampão de perfusão quente a uma velocidade de bomba de três mililitros por minuto.

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