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Diabetologia. 2016 Nov;59(11):2387-2392. doi: 10.1007/s00125-016-4049-6. Epub 2016 Jul 21.

Sequential intravital imaging reveals in vivo dynamics of pancreatic tissue transplanted under the kidney capsule in mice.

Author information

1
Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.
2
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands.
3
The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, L1, Stockholm, Sweden.
4
Department of Tumor Immunology, Radboudumc, Nijmegen, the Netherlands.
5
Institute of Molecular Health Sciences, ETH Zürich, Zürich, Switzerland.
6
Cancer Genomics Center, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, the Netherlands.
7
Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands. e.koning@hubrecht.eu.
8
Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands. e.koning@hubrecht.eu.

Abstract

AIMS/HYPOTHESIS:

Dynamic processes in pancreatic tissue are difficult to study. We aimed to develop an intravital imaging method to longitudinally examine engraftment, vascularisation, expansion and differentiation in mature islets or embryonic pancreases transplanted under the kidney capsule.

METHODS:

Isolated pancreatic islets from adult mice and murine embryonic day (E)12.5 pancreases containing fluorescent biomarkers were transplanted under the kidney capsule of immunodeficient recipient mice. Human islet cells were dispersed, transduced with a lentivirus expressing a fluorescent label and reaggregated before transplantation. Graft-containing kidneys were positioned subcutaneously and an imaging window was fitted into the skin on top of the kidney. Intravital imaging using multiphoton microscopy was performed for up to 2 weeks. Volumes of fluorescently labelled cells were determined as a measure of development and survival.

RESULTS:

Transplanted islets and embryonic pancreases showed good engraftment and remained viable. Engraftment and vascularisation could be longitudinally examined in murine and human islet cells. Murine islet beta cell volume was unchanged over time. Transplanted embryonic pancreases increased to up to 6.1 times of their original volume and beta cell volume increased 90 times during 2 weeks.

CONCLUSIONS/INTERPRETATION:

This method allows for repeated intravital imaging of grafts containing various sources of pancreatic tissue transplanted under the kidney capsule. Using fluorescent markers, dynamic information concerning engraftment or differentiation can be visualised and measured.

KEYWORDS:

Development; Embryonic; Human; Imaging; Intravital; Islets; Microscopy; Mouse; Pancreas; Transplantation

PMID:
27443307
PMCID:
PMC5506096
DOI:
10.1007/s00125-016-4049-6
[Indexed for MEDLINE]
Free PMC Article

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