Pancreatic Islet Viability Assessment Using Hyperspectral Imaging of Autofluorescence

Cells. 2023 Sep 19;12(18):2302. doi: 10.3390/cells12182302.

Abstract

Islets prepared for transplantation into type 1 diabetes patients are exposed to compromising intrinsic and extrinsic factors that contribute to early graft failure, necessitating repeated islet infusions for clinical insulin independence. A lack of reliable pre-transplant measures to determine islet viability severely limits the success of islet transplantation and will limit future beta cell replacement strategies. We applied hyperspectral fluorescent microscopy to determine whether we could non-invasively detect islet damage induced by oxidative stress, hypoxia, cytokine injury, and warm ischaemia, and so predict transplant outcomes in a mouse model. In assessing islet spectral signals for NAD(P)H, flavins, collagen-I, and cytochrome-C in intact islets, we distinguished islets compromised by oxidative stress (ROS) (AUC = 1.00), hypoxia (AUC = 0.69), cytokine exposure (AUC = 0.94), and warm ischaemia (AUC = 0.94) compared to islets harvested from pristine anaesthetised heart-beating mouse donors. Significantly, with unsupervised assessment we defined an autofluorescent score for ischaemic islets that accurately predicted the restoration of glucose control in diabetic recipients following transplantation. Similar results were obtained for islet single cell suspensions, suggesting translational utility in the context of emerging beta cell replacement strategies. These data show that the pre-transplant hyperspectral imaging of islet autofluorescence has promise for predicting islet viability and transplant success.

Keywords: autofluorescence; hyperspectral; islet; multispectral; transplantation; viability.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytokines
  • Humans
  • Hyperspectral Imaging
  • Hypoxia
  • Insulin-Secreting Cells*
  • Islets of Langerhans* / diagnostic imaging
  • Mice

Substances

  • Cytokines

Grants and funding

This research was supported by the Juvenile Diabetes Research Foundation through the JDRF award 1-INO-2019-788-S-B to S.T.G and E.G. and partially supported by a Discovery Project grant from the Australian Research Council (DP170101863) and the Centre of Excellence scheme (CE140100003), as well as the UNSW SHARP scheme to E.G. The research was also supported by National Health Medical Research Council (NHMRC) grants (GNT1130222; GNT1146493; GNT1189235) and the Twin Towns Services Community Foundation to S.T.G. S.T.G. is a NHMRC Senior Research Fellow (GNT1140691).