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Author Spotlight: Techniques and Challenges in Islet Purification for Xenotransplantation
Published on: June 7, 2024
Pancreas preservation with amphotericin B deteriorates islet yield for porcine islet isolation
Sayaka Sawada1, Chika Miyagi-Shiohira1, Kazuho Kuwae1
1Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, Nishihara, Japan.
Background:
Amphotericin B is a crucial agent in the management of serious systemic fungal infections. It is also known to be cytotoxic. In this study, we evaluated the effect of amphotericin B added to the preservation solution on islet yield during islet isolation.
Methods:
Porcine pancreata were preserved in the preservation solution with or without amphotericin B (0.25 μg/mL) for approximately 18 hours at 4°C, and then islet isolation was performed. An optimized number (1750 IE) of isolated islets from each group were transplanted into streptozotocin-induced diabetic mice. The culture of isolated islets and acinar tissue with amphotericin B was also evaluated.
Results:
The islet yield before and after purification in the amphotericin B (-) group was significantly higher than that in the amphotericin B (+) group. After islet transplantation into diabetic mice, blood glucose levels reached the normoglycemic range, with 50% and 0% of that of the diabetic mice in the amphotericin B (-) and amphotericin B (+) groups, respectively. In the culture study, amphotericin B was found to be cytotoxic to porcine islets and acinar tissue.
Conclusions:
Amphotericin B added to the preservation solution deteriorates islet yield during porcine islet isolation. Thus, the use of amphotericin B should be considered carefully for the preservation of the pancreas for islet isolation and islet culture before islet transplantation.
Insights
Adding amphotericin B to preservation solutions significantly reduces islet yield during porcine islet isolation. This antifungal agent is cytotoxic, negatively impacting islet isolation and transplantation success.
Area of Science:
- Transplantation Biology
- Islet Isolation
- Fungal Infection Management
Background:
- Amphotericin B is vital for treating severe fungal infections.
- Amphotericin B exhibits known cytotoxic properties.
- The study investigates Amphotericin B's impact on islet isolation.
Purpose of the Study:
- To evaluate the effect of Amphotericin B in preservation solutions on islet yield.
- To assess the impact of Amphotericin B on islet isolation and islet culture.
- To determine the safety and efficacy of using Amphotericin B during pancreatic preservation for islet isolation.
Main Methods:
- Porcine pancreata were preserved with or without Amphotericin B (0.25 μg/mL) for 18 hours at 4°C.
- Islet isolation was performed, and optimized islet numbers (1750 IE) were transplanted into diabetic mice.
- The cytotoxicity of Amphotericin B on isolated islets and acinar tissue was assessed in culture.
Main Results:
- Islet yield was significantly higher in the absence of Amphotericin B.
- Transplanted islets from the Amphotericin B-free group normalized blood glucose in diabetic mice (50% efficacy).
- Amphotericin B demonstrated cytotoxicity to porcine islets and acinar tissue in vitro.
Conclusions:
- Amphotericin B in preservation solutions impairs islet yield during porcine islet isolation.
- Careful consideration of Amphotericin B use is recommended for pancreas preservation for islet isolation and culture.
- The cytotoxic nature of Amphotericin B necessitates caution before islet transplantation.
