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Optimizing post-transplantation detection of subcutaneously transplanted islets using dithizone staining.

James Lu¹, Matthew Ishahak¹, Marlie M Maestas¹

¹Division of Endocrinology, Metabolism and Lipid Research, Washington University School of Medicine, St. Louis, MO USA.

|May 20, 2026

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Dithizone staining effectively labels transplanted pancreatic islet grafts in mice, enabling precise localization and recovery. This method enhances subcutaneous islet transplantation research by providing a reliable tool for graft identification and analysis.

Area of Science:

Endocrinology
Cell Biology
Transplantation Science

Background:

Pancreatic islet transplantation aims to restore insulin independence in type 1 diabetes.
Human pluripotent stem cell-derived islets show clinical promise but face challenges in site optimization and graft localization.
The subcutaneous space is an accessible transplantation site, yet its utility is limited by poor vascularization and difficulty in locating grafts.

Purpose of the Study:

To investigate the efficacy of dithizone staining for reliable localization of transplanted islet grafts within the subcutaneous space.
To establish a standardized framework for evaluating subcutaneous islet transplantation using dithizone and automated islet quantification.

Main Methods:

Stem cell-derived islets (SC-islets) and primary human islets were transplanted into immunodeficient mice.

Keywords:

Diabetes Dithizone staining Islet transplantation SC-islets

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Automated islet equivalent quantification standardized islet volume prior to transplantation.

Graft function was monitored via human C-peptide levels.

Dithizone staining was applied to harvested tissue for graft localization, followed by immunohistochemical validation.

Main Results:

Automated quantification ensured consistent islet dosing.
Transplanted islets maintained function, evidenced by detectable human C-peptide.
Dithizone staining clearly and selectively labeled insulin-producing β-cell grafts, enabling reliable localization and recovery.
Immunohistochemistry confirmed insulin-expressing cells in isolated grafts.

Conclusions:

Dithizone staining offers a rapid, cost-effective method for identifying subcutaneous islet grafts, overcoming a key research limitation.
This technique, combined with standardized islet quantification, provides a reproducible framework for subcutaneous islet transplantation studies.
Further research should explore dithizone's effects and applicability to other sites and imaging methods.