Canine Beta Cell Line Generation via Scid Mouse Grafting
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for treating canine diabetes are limited by the lack of reliable and reproducible methods for generating functional canine beta cell lines, which are essential for cell therapy and drug screening, due to the underdeveloped understanding of canine pancreas development and the inability to expand cryopreserved Islets of Langerhans.
Innovation Solution
A method involving the grafting of immature canine pancreatic tissue into scid mice, followed by sub-grafting and enrichment steps, to generate stable and functional canine beta cell lines that can be amplified for clinical and commercial use, using lentiviral vectors and specific promoters to immortalize and de-immortalize cells as needed.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If cryopreserved Islets of Langerhans are used for cell therapy, then the treatment can be applied to diabetic animals, but the amount of islets available is strictly limited by the amount collected on each organ and cannot be expanded
Solution Approach 1:
The patent segments the pancreas into distinct functional regions, specifically isolating the endocrine islets from the exocrine tissue. This segmentation allows for the selective collection and expansion of beta cells while leaving the bulk of the pancreatic tissue unused, thereby increasing the effective quantity of expandable cells available for therapy.
Solution Approach 2:
The patent performs preliminary enzymatic digestion and mechanical dissociation of the pancreatic tissue before cell collection. This preliminary processing step breaks down the tissue matrix and releases individual cells, enabling subsequent expansion in culture media. Without this preliminary action, the islets would remain intact and non-expandable.
2Reliability
If daily insulin injections are administered for diabetes treatment, then the treatment is effective for all types of diabetes in dogs, but it represents a significant financial burden and results in significant deterioration in quality of life
Solution Approach 1:
The patent generates autologous beta cell lines from the patient's own pancreatic tissue, creating a self-sufficient cell therapy system. The derived cell lines can be expanded and used to produce insulin or transplanted to restore endogenous insulin production, eliminating the need for continuous external insulin administration and reducing the financial and quality-of-life burden on patients.
3Productivity
If canine beta cell lines are generated from immature pancreas, then functional cells can be produced for therapy and drug screening, but the method requires complex grafting procedures into scid mice and multiple enrichment steps
Solution Approach 1:
The patent uses scid mice as intermediary hosts to facilitate the generation of canine beta cell lines. The immunodeficient mice provide a permissive environment where canine pancreatic cells can be grafted, enriched, and expanded without rejection. This intermediary system simplifies the overall process by providing a controlled platform for cell manipulation that would otherwise be impossible in vivo.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the production of homogenous, stable, and functional canine beta cell lines capable of producing insulin, enabling effective cell therapy and diagnosis of canine diabetes, as well as the screening of new drugs for diabetes treatment.
Implementation Method 1
transducing and co-transducing immature canine pancreas cells with i) a lentiviral vector expressing SV40 LargeT antigen under the control of the insulin promoter, or ii) with a lentiviral vector expressing SV40 LargeT antigen under the control of the insulin promoter and a lentiviral vector expressing hTert under the control of the insulin promoter
Implementation Method 2
allowing the transduced immature pancreas cells to develop insulinoma-like structures, wherein the immature canine pancreases cells in insulinoma-like structures have differentiated to insulin-producing pancreatic beta cells
Implementation Method 3
The pancreatic tissue was treated with collagenase XI (1 mg/ml RPMI) (Sigma-Aldrich) during 30 minutes at 37° C.
Data Source
AI summary
The present invention relates to a method for preparing commercial scale quantities of canine functional beta cells and to the establishment of cell lines from immature canine pancreatic tissues. It also relates to a method of diagnosis using canine beta cell tumours or cells derived thereof. The method comprises sub-transplantation procedure to enrich the graft in proliferating beta cells, allowing generating canine Beta cell lines. Such lines express, produce and secrete insulin upon glucose stimulation.