The great demand for a cure that can finally overcome diabetes type 1, restoring normo-glycemic levels in patients, has acquired interest in research. Nowadays the transplantation of pancreatic islets in patients with type 1 diabetes resulted a promising therapy able to manage glucose homeostasis. However, this treatment remains an experimental therapy limited to the most severe cases of diabetes. One of the main drawbacks is the graft failure in the early post-transplant period that can be due to a non-optimal condition of the transplanted islets. This issue is related to the lack of reliable assessment methods of islet functionality pre-transplantation. Currently the gold standard used for the assessment of the beta cells functionality is the in vitro glucose stimulated insulin release (GSIR) assay which is a time-consuming test in which pancreatic islets are incubated in direct contact with the stimulating glucose solutions. The present thesis aims to implement a microfluidic device as a platform to assess pancreatic islets functionality. This platform, called GSIR-on-a-chip, has as main goal the study of islets functionality in a more physiological-like environment. Indeed, the islets in the chip are embedded in a hydrogel that reproduces the diffusion-driven mass transport of the solutes (as glucose and insulin), allowing a more physiological response. In addition to that, the microfluidic platform, compared to the standard GSIR, minimizes the duration of the whole assay, the number of islets and the volumes of reagents needed. This microfluidic platform has demonstrated to assess cell functionality in an efficient manner, taking the transplantation of pancreatic islets a step closer to the widespread use as therapy for type 1 diabetes.
Il trapianto di isole pancreatiche è una terapia sperimentale che ad oggi ha mostrato risultati promettenti nella ricerca di una cura al Diabete di tipo 1. Tale terapia rimane però consigliata ad una tipologia ristretta di pazienti con gravi complicanze poiché non garantisce una efficacia del trapianto a lungo termine. Le isole pancreatiche trapiantate perdono infatti la loro funzionalità post-trapianto, e questa problematica spesso è dovuta alla mancanza di metodi efficienti per la valutazione della funzionalità delle isole pre-trapianto. Al momento il gold standard utilizzato per valutare la funzionalità in vitro è il saggio Glucose Stimulated Insulin Release (GSIR). Tale test risulta essere dispendioso in termini di tempo, numero di isole e reagenti utilizzati. Al fine di trovare una procedura migliore, il presente progetto di tesi mira ad ottimizzare una piattaforma microfluidica capace di valutare la funzionalità delle isole pancreatiche in minor tempo e con un limitato dispendio di isole pancreatiche e reagenti. L’obiettivo principale del GSIR-on-a-chip è quello di testare le isole in un ambiente più simile a quello fisiologico, capace di ricreare la componente diffusiva del processo di trasporto di massa dei soluti quali glucosio ed insulina, permettendo quindi una migliore risposta cellulare. In generale il GSIR-on-a-chip ha dimostrato di poter testare la funzionalità delle isole in maniera efficiente e con risultati comparabili al GSIR standard rendendo dunque il trapianto di isole pancreatiche una terapia sempre più concreta e diffusa per la cura del diabete.
GSIR-on-a-chip : optimization of an in vitro microfluidic platform to assess pancreatic islet functionality
Recchione, Lorenza
2020/2021
Abstract
The great demand for a cure that can finally overcome diabetes type 1, restoring normo-glycemic levels in patients, has acquired interest in research. Nowadays the transplantation of pancreatic islets in patients with type 1 diabetes resulted a promising therapy able to manage glucose homeostasis. However, this treatment remains an experimental therapy limited to the most severe cases of diabetes. One of the main drawbacks is the graft failure in the early post-transplant period that can be due to a non-optimal condition of the transplanted islets. This issue is related to the lack of reliable assessment methods of islet functionality pre-transplantation. Currently the gold standard used for the assessment of the beta cells functionality is the in vitro glucose stimulated insulin release (GSIR) assay which is a time-consuming test in which pancreatic islets are incubated in direct contact with the stimulating glucose solutions. The present thesis aims to implement a microfluidic device as a platform to assess pancreatic islets functionality. This platform, called GSIR-on-a-chip, has as main goal the study of islets functionality in a more physiological-like environment. Indeed, the islets in the chip are embedded in a hydrogel that reproduces the diffusion-driven mass transport of the solutes (as glucose and insulin), allowing a more physiological response. In addition to that, the microfluidic platform, compared to the standard GSIR, minimizes the duration of the whole assay, the number of islets and the volumes of reagents needed. This microfluidic platform has demonstrated to assess cell functionality in an efficient manner, taking the transplantation of pancreatic islets a step closer to the widespread use as therapy for type 1 diabetes.| File | Dimensione | Formato | |
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https://hdl.handle.net/10589/187195