title: Sequential intravital imaging reveals in vivo dynamics of pancreatic tissue transplanted under the kidney capsule in mice.
abstract: 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.
  
  
  
  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.
  
  
  
  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.
  
  
  
  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.
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.
authors.forename: Léon
authors.initials: L
authors.lastname: van Gurp
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.|Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Cindy J M
authors.initials: CJM
authors.lastname: Loomans
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.|The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, L1, Stockholm, Sweden.
authors.forename: Pim P
authors.initials: PP
authors.lastname: van Krieken
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.
authors.forename: Gitanjali
authors.initials: G
authors.lastname: Dharmadhikari
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.|Department of Tumor Immunology, Radboudumc, Nijmegen, the Netherlands.
authors.forename: Erik
authors.initials: E
authors.lastname: Jansen
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands.|Institute of Molecular Health Sciences, ETH Zürich, Zürich, Switzerland.
authors.forename: Femke C A S
authors.initials: FCAS
authors.lastname: Ringnalda
authors.affiliation: Cancer Genomics Center, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, the Netherlands.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Beerling
authors.affiliation: Cancer Genomics Center, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, the Netherlands.
authors.forename: Jacco
authors.initials: J
authors.lastname: van Rheenen
authors.affiliation: Hubrecht Institute-KNAW and University Medical Center Utrecht, Uppsalalaan 8, 3584CT, Utrecht, the Netherlands. e.koning@hubrecht.eu.|Department of Internal Medicine, Leiden University Medical Center, Leiden, the Netherlands. e.koning@hubrecht.eu.
authors.forename: Eelco J P
authors.initials: EJP
authors.lastname: de Koning
mesh_terms: D000818:Animals; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007515:Islets of Langerhans / Q000166:cytology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D010179:Pancreas / Q000166:cytology* / Q000196:embryology* / Q000378:metabolism; D011247:Pregnancy
keywords: Development; Embryonic; Human; Imaging; Intravital; Islets; Microscopy; Mouse; Pancreas; Transplantation