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4-Dimensional Intravital Microscopy: A New Model for Studies of Leukocyte Recruitment and Migration in Hepatocellular Cancer in Mice

  • Original Article
  • Published:
Journal of Gastrointestinal Surgery

Abstract

Introduction

Although it is accepted that the immune system plays a role in the prognosis of hepatocellular carcinoma (HCC), the exact mechanisms of leukocyte recruitment into HCC are poorly understood. Progress in the study of this aspect has been hindered by technical limitations.

Materials and Methods

In the present study, we describe the use of 4D intravital microscopy which represents an advantageous technology for the investigation of the microvascular system and leukocyte migration in HCC. To establish 4D intravital microscopy, we used a HCC tumor model in transgenic mice expressing enhanced green fluorescent protein in specific leukocyte subpopulations and combined digital time-lapse recording, laser scanning confocal microscopy, and 3D reconstruction. Using this technology, we studied the intra- and extravascular leukocyte adhesion and migration in HCC in vivo at the single-cell level.

Results

We showed that although vessel density in HCC was lower than in normal liver, tumor tissue was moderately infiltrated with leukocytes of lymphoid and myeloid origin. Most tumor-infiltrating leukocytes migrated in a random manner frequently changing direction of migration in the tumor tissue. The migration velocity of myeloid and lymphoid leukocytes in HCC tissue was not different.

Discussion

These results demonstrated that 4D intravital microscopy has potential to be a powerful tool in the study of mechanisms of leukocyte recruitment and intratumoral migration in HCC.

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Abbreviations

HCC:

Hepatocellular carcinoma

EGFP:

Enhanced green fluorescent protein

4D:

4-Dimensional

TRITC:

Tetramethylrhodamine isothiocyanate

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Acknowledgment

We thank Dr. U. Engel and Dr. Ch. Ackermann (Nikon Imaging Center, University of Heidelberg) for technical support in microscopy. We thank C. Bernardi for her excellent assistance. We thank Dr T. Graf (Albert Einstein College of Medicine, New York, USA) for providing of lys-EGFP-ki mice. We thank Prof. K. Ley (University of Virginia, Charlottesville, USA) for providing CD2-EGFP mice.

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Authors and Affiliations

  1. Department of Surgery, University of Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany

    Takayuki Takeichi, Paulius Mocevicius, Jan Schmidt & Eduard Ryschich

  2. Department of Pediatrics, University of Heidelberg, Im Neuenheimer Feld 110, 69120, Heidelberg, Germany

    Guido Engelmann

  3. Department of Transplantation and Pediatric Surgery, Postgraduate School of Medical Science, Kumamoto University, 1-1-1 Honjyo, Kumamoto, 860-8556, Japan

    Takayuki Takeichi

Authors
  1. Takayuki Takeichi
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  2. Guido Engelmann
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  3. Paulius Mocevicius
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  4. Jan Schmidt
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  5. Eduard Ryschich
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Corresponding author

Correspondence to Takayuki Takeichi.

Electronic Supplementary Materials

Below is the link to the electronic supplementary material.

Representative time-lapse movies of leukocyte migration in HCC in Lys-EGFP-ki. Leukocyte migration was recorded 30 min and was compressed to 10 s in time-lapse mode. (AVI 20833 kb)

Representative time-lapse movies of leukocyte migration in CD2-EGFP+ mice. Leukocyte migration was recorded 30 min and was compressed to 10 s in time-lapse mode. (AVI 17642 kb)

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Takeichi, T., Engelmann, G., Mocevicius, P. et al. 4-Dimensional Intravital Microscopy: A New Model for Studies of Leukocyte Recruitment and Migration in Hepatocellular Cancer in Mice. J Gastrointest Surg 14, 867–872 (2010). https://doi.org/10.1007/s11605-010-1179-x

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  • DOI: https://doi.org/10.1007/s11605-010-1179-x

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