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Diabetes 54:2287-2293, 2005
© 2005 by the American Diabetes Association, Inc.

Donor Islet Endothelial Cells Participate in Formation of Functional Vessels Within Pancreatic Islet Grafts

Daniel Nyqvist1, Martin Köhler1, Helene Wahlstedt1, and Per-Olof Berggren1,2

1 The Rolf Luft Center for Diabetes Research, Department of Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, Stockholm, Sweden
2 Diabetes Research Institute, University of Miami School of Medicine, Miami, Florida

Pancreatic islet transplantation has emerged as a therapy for type 1 diabetes and is today performed using both freshly isolated and cultured islets. Islet blood vessels are disrupted during islet isolation; therefore, proper revascularization of the transplanted islets is of great importance for islet graft function and survival. We have studied intraislet endothelial cells after islet isolation, during islet culture, and following islet transplantation. By isolating islets from the transgenic Tie2-GFP (green fluorescent protein) mouse, characterized by an endothelial cell–specific expression of GFP, living endothelial cells could be studied in intact islets utilizing two-photon laser-scanning microscopy (TPLSM). Intraislet endothelial cells were found to survive islet transplantation but to rapidly disappear during islet culture. By transplanting freshly isolated Tie2-GFP islets and applying a novel ex vivo model for simultaneous perfusion and TPLSM imaging of the graft-bearing kidneys, GFP fluorescent endothelial cells were found to extensively contribute to vessels within the islet graft vasculature. Real-time imaging of the flow through the islet graft vasculature confirmed that the donor-derived vessels were functionally integrated. Hence, intraislet endothelial cells have the capability of participating in revascularization of pancreatic islets subsequent to transplantation. Therefore, preservation of intraislet endothelial cell mass may improve long-term graft function.

Address correspondence and reprint requests to Daniel Nyqvist, The Rolf Luft Center for Diabetes Research L3, Department of Molecular Medicine, Karolinska Institutet, Karolinska University Hospital Solna, S-17176 Stockholm, Sweden. E-mail: daniel.nyqvist{at}molmed.ki.se

Abbreviations: CLSM, confocal laser-scanning microscopy; ECGS, endothelial cell growth supplement; FGF, fibroblast growth factor; FITC, fluoroscein isothiocyanate; GRP, green fluorescent protein; TPLSM, two-photon laser-scanning microscopy; TRITC, tetrarhodamine isothiocyanate; VEGF, vascular endothelial growth factor


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