Quantification of Unmethylated Insulin DNA Using Methylation Sensitive Restriction Enzyme Digital Polymerase Chain Reaction
Transplant International, vol. 35, pp. 10167
Abstract
Assessment of specific β-cell death can be used to determine the quality and viability of pancreatic islets prior to transplantation and hence predict the suitability of the pancreas for isolation. Recently, several groups have demonstrated that unmethylated insulin (INS)-DNA is correlated to β-cell death in type 1 diabetes patients and during clinical islet isolation and subsequent transplantation. Here, we present a step-by-step protocol of our novel developed method for quantification of the relative amount of unmethylated INS-DNA using methylation sensitive restriction enzyme digital polymerase chain reaction This method provides a novel and sensitive way to quantify the relative amount of β-cell derived unmethylated INS-DNA in cellular lysate. We therefore suggest that this technique can be of value to reliably determine the purity of an islet preparation and may also serve as a measure of the quality of islets prior to transplantation measuring unmethylated INS-DNA as a reflection of the relative amount of lysed β-cells.
Authors 9
-
Fenna E.M. van de Leemkolk Aachen Department of Surgery, Leiden University Medical Center LUMC Transplant Center
Leiden University Medical Center
Affiliation as printed
Department of Surgery, Leiden University Medical Center, Leiden, Netherlands
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Department of Surgery, Netherlands
LUMC Transplant Center, Netherlands
-
Leiden University Medical Center
Affiliation as printed
Department of Ophthalmology, Leiden University Medical Center, Leiden, Netherlands
Department of Ophthalmology, Netherlands
-
Leiden University Medical Center
Affiliation as printed
Department of Ophthalmology, Leiden University Medical Center, Leiden, Netherlands
Department of Ophthalmology, Netherlands
-
Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, Netherlands
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Department of Internal Medicine, Netherlands
LUMC Transplant Center, Netherlands
-
Volkert A.L. Huurman Aachen Department of Surgery, Leiden University Medical Center LUMC Transplant Center
Leiden University Medical Center
Affiliation as printed
Department of Surgery, Leiden University Medical Center, Leiden, Netherlands
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Department of Surgery, Netherlands
LUMC Transplant Center, Netherlands
-
Ian P.J. Alwayn Aachen Department of Surgery, Leiden University Medical Center LUMC Transplant Center
Leiden University Medical Center
Affiliation as printed
Department of Surgery, Leiden University Medical Center, Leiden, Netherlands
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Department of Surgery, Netherlands
LUMC Transplant Center, Netherlands
-
Leiden University Medical Center · University of Oxford
Affiliation as printed
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Nuffield Department of Surgical Sciences, University of Oxford, Oxford, United Kingdom
LUMC Transplant Center, Netherlands
Nuffield Department of Surgical Sciences, United Kingdom
-
Leiden University Medical Center
Affiliation as printed
Department of Ophthalmology, Leiden University Medical Center, Leiden, Netherlands
Department of Ophthalmology, Netherlands
-
Leiden University Medical Center
Affiliation as printed
Department of Internal Medicine, Leiden University Medical Center, Leiden, Netherlands
LUMC Transplant Center, Leiden University Medical Center, Leiden, Netherlands
Department of Internal Medicine, Netherlands
LUMC Transplant Center, Netherlands
Cited by 4 stored of 5
4 results
No patents citing this paper on Lens.org (checked 2026-10-11).