Poster Preview
Type 1 Diabetes (T1D) is an autoimmune disease characterized by the inflammation and destruction of pancreatic islets by infiltrating immune cells. As T1D incidence continues to surge worldwide, it is necessary to develop therapies capable of strengthening and protecting islet function.
College
College of Arts & Sciences
Mentor Information
Brant R. Burkhardt
Description
Type 1 Diabetes (T1D) is an autoimmune disease characterized by the inflammation and destruction of pancreatic islets by infiltrating immune cells. As T1D incidence continues to surge worldwide, it is necessary to develop therapies capable of strengthening and protecting islet function.
Our prior research has previously demonstrated the protective antioxidant effects of Cornus officinalis (CO), a medicinal herb with known anti-diabetes properties, using 1.1B4 and NIT-1 transformed pancreatic β-cell lines. Although these cell lines are valuable and cost-efficient β-cell models, they are not fully representative of primary β-cell biology, which may affect their antioxidant response to CO.
To this end, we have expanded to using murine islets as a more biologically relevant model to evaluate the cytoprotective effects of CO. However, isolation of murine pancreatic islets is a technically challenging and time-consuming procedure often resulting in poor yield or quality. Therefore, we sought to enhance the islet isolation protocol for islet yield and viability. To achieve this, islets were harvested from normoglycemic female nonobese diabetic (NOD) mice, which are validated models of T1D. Murine pancreases were extracted and digested with collagenase P, then purified and isolated using a novel double-filtration system.
This enhanced procedure resulted in increased islet viability and a robust yield of 250-600 islets per pancreas. In addition, viability and insulin secretion assays revealed these islets to be highly functional and insulinogenic. These findings suggest that our recently developed filtration method is highly efficient, facilitating the utilization of islets for T1D studies.
Protocol for NOD Islet Isolation Using a Novel Two-Filter Purification System
Type 1 Diabetes (T1D) is an autoimmune disease characterized by the inflammation and destruction of pancreatic islets by infiltrating immune cells. As T1D incidence continues to surge worldwide, it is necessary to develop therapies capable of strengthening and protecting islet function.
Our prior research has previously demonstrated the protective antioxidant effects of Cornus officinalis (CO), a medicinal herb with known anti-diabetes properties, using 1.1B4 and NIT-1 transformed pancreatic β-cell lines. Although these cell lines are valuable and cost-efficient β-cell models, they are not fully representative of primary β-cell biology, which may affect their antioxidant response to CO.
To this end, we have expanded to using murine islets as a more biologically relevant model to evaluate the cytoprotective effects of CO. However, isolation of murine pancreatic islets is a technically challenging and time-consuming procedure often resulting in poor yield or quality. Therefore, we sought to enhance the islet isolation protocol for islet yield and viability. To achieve this, islets were harvested from normoglycemic female nonobese diabetic (NOD) mice, which are validated models of T1D. Murine pancreases were extracted and digested with collagenase P, then purified and isolated using a novel double-filtration system.
This enhanced procedure resulted in increased islet viability and a robust yield of 250-600 islets per pancreas. In addition, viability and insulin secretion assays revealed these islets to be highly functional and insulinogenic. These findings suggest that our recently developed filtration method is highly efficient, facilitating the utilization of islets for T1D studies.
