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マウスにおける肝部分切除後の再生肝細胞の単離
マウスにおける肝部分切除後の再生肝細胞の単離
JoVE Journal
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JoVE Journal Medicine
Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice

マウスにおける肝部分切除後の再生肝細胞の単離

Full Text
5,809 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.

脂質を含んだ肝細胞は肝臓の再生に固有のものですが、通常は密度勾配遠心分離で失われます。ここでは、脂肪肝細胞を保持する最適化された細胞分離プロトコルを提示し、マウスの部分肝切除後に再生肝細胞の代表的な集団を生み出します。

このプロトコルの全体的な目標は、密度勾配遠心分離を必要とせずに、部分肝切除後のマウスにおける肝細胞単離のための最適化された方法を提示することです。通常、初期の再生中に脂質を含んだ肝細胞の大部分が見られます。低密度のために密度勾配遠心分離で通常失われるこれらの脂肪肝細胞は、このプロトコルで保持されます。

灌流装置を準備するには、ルアーロックコネクタを使用して、26ゲージのIUIカニューレをチューブの出口端に接続します。次に、チューブを洗い流し、チューブの入口端を水浴中の予熱した灌流バッファーチューブに挿入します。毎分3ミリリットルのポンプ速度で温かい灌流バッファーでプライミングします。

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