Pancreatic Islet Isolation: From the Mouse to the Clinic by Miriam Ramírez-Domínguez

By Miriam Ramírez-Domínguez

Pancreatic islets make up the endocrine pancreas they usually include the single resource of insulin within the physique, beta cells. for that reason, entry to top of the range arrangements of pancreatic islets is prime for in vitro experiences and to check pre-clinical functions in animal versions in vivo. entry to fit human islets is usually an important to enhance transplantation techniques for diabetes. Given the susceptibility of pancreatic islets to the enzymatic digestion and mechanical tension required to procure them, the isolation of islets is frequently regarded as the fragile “work of a craftsman”. This e-book, that's aimed toward rookies and specialists alike, is a survey of the present state-of–the-art during this box and it centres at the demanding situations, pitfalls and peculiarities of pancreatic islet isolation within the diversified species utilized in pre-clinical and medical functions. It explores the similarities and changes among human islets and people from different appropriate species (rodents, pigs and non-human primates), and the way those impact islet isolation. the final word target of this publication is to enhance the result of islet isolation and transplantation in pre-clinical and scientific applications.

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36. Gray D, Sutton R, McShane P, Peters M, Morris P. Exocrine contamination impairs implantation of pancreatic islets transplanted beneath the kidney capsule. J Surg Res. 1988;45(5):432–42. 37. Loganathan G, Dawra RK, Pugazhenthhi S, Guo Z, Soltani SM, Wiseman A, Sanders MA, Papas KK, Velayutham K, Saluja AK, Sutherland DE, Hering BJ, Balamurugan AN. Insulin degradation by acinar cell proteases creates a dysfunctional environment for human islets before/after transplantation: benefits for α-1 antytripsin treatment.

The aim of pancreatic islet isolation is obtaining viable, pure and functional islets, either for in vitro or in vivo studies (Fig. 1). However, to obtain a successful yield and good quality islets, different key aspects must be taken into account: the type and concentration of the digestive enzyme, the method of enzyme administration, the temperature and duration of the pancreas digestion, the method for islet purification and the culture conditions following isolation [6]. It Fig. 1 Fluorescence micrograph showing the overall viability of the islets by FDA/PI vital staining.

The most common islet culture medium for murine islets is RPMI 1640 (Rosewell Park Memorial Institute Medium). However, some laboratories prefer to use CMRL 1066 (Connaught Medical Research Laboratories), which is also widely used to culture human pancreatic islets. Preference for CMRL 1066 is based on the observation that some immune cells, such as dendritic and endothelial cells, cannot grow in that medium, which may therefore induce a decrease in alloreactivity. One component of the medium that is key for the physiology of the islets is glucose.

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