Facile mechanical shaking method is an improved isolation approach for islet preparation and transplantation

Abstract

Successful islet isolation is crucial for islet transplantation and cell treatment for type 1 diabetes. Current isolation methods are able to obtain 500–1,000 islets per rat, which results in a waste of ≥50% of total islets. In the present study, a facile mechanical shaking method for improving islet yield (up to 1,500 per rat) was developed and summarized, which was demonstrated to be more effective than the existing well-established stationary method. The present results showed that isolated islets have a maximum yield of 1,326±152 when shaking for 15 min for the fully-cannulated pancreas. For both fully-cannulated and half-cannulated pancreas in the presence of rat DNAse inhibitor, the optimal shaking time was amended to 20 min with a further increased yield of 1,344±134 and 1,286±124 islets, respectively. Furthermore, the majority of the isolated islets were morphologically intact with a well-defined surface and almost no central necrotic zone, which suggested that the condition of islets obtained via the mechanical shaking method was consistent with the stationary method. Islet size distribution was also calculated and it was demonstrated that islets from the stationary method exhibited the same size distribution as the non-cannulated group, which had more larger islets than the fully-cannulated and half-cannulated groups isolated via the shaking method. In addition, the results of glucose challenge showed that the refraction index of each group was >2.5, which indicated the well-preserved function of isolated islets. Furthermore, the transplanted islets exhibited a therapeutic effect after 1 day of transplantation; however, they failed to control blood glucose levels after ~7 days of transplantation. In conclusion, these results demonstrated that the facile mechanical shaking method may markedly improve the yield of rat islet isolation, and in vitro and in vivo investigation demonstrated the well-preserved function of isolated islets in the control of blood glucose. Therefore, the facile mechanical shaking method may be an alternative improved procedure to obtain higher islet yield for islet preparation and transplantation in the treatment of type 1 diabetes.

Human Pluripotent Stem Cells: A Unique Tool for Toxicity Testing in Pancreatic Progenitor and Endocrine Cells

“…Interestingly, islets isolated from juvenile porcine pancreata have greater expression of b-cell-related genes compared to those isolated from adult pigs (121), but are functionally immature and require in vitro maturation following isolation (122)(123)(124). Further, it is possible to isolate up to 5,000 islets/g juvenile porcine pancreas (122), compared to a typical yield of~200-400 islets total per mouse pancreas and 600-800 islets per rat pancreas depending on the strain (125)(126)(127). Thus, pig islets may be a useful tool for toxicity screening, although species differences will always remain a concern for translation.…”

Recovery of isolated pancreatic isletsderived from gut leak-induced diabetes type ii mouse model

“…The techniques for pancreas digestion can be done by mainly two ways, perfusion and mechanical with the digestion solution (Yin et al, 2016). The advantages of the isolating islets by using perfusion technique are allowing the digestion enzyme perfuse to every corner of the pancreas anatomically and minimize the wound of the islets (Carter et al, 2009).…”

“…The advantages of the isolating islets by using perfusion technique are allowing the digestion enzyme perfuse to every corner of the pancreas anatomically and minimize the wound of the islets (Carter et al, 2009). However, this technique must be done in the right process to get higher yield of the islets (Yin et al, 2016).…”