Abstract
Rapid, efficient, and economic method for the isolation and purification of islets has been pursued by numerous islet-related researchers. In this study, we compared the advantages and disadvantages of our developed patented method with those of commonly used conventional methods (Ficoll-400, 1077, and handpicking methods). Cell viability was assayed using Trypan blue, cell purity and yield were assayed using diphenylthiocarbazone, and islet function was assayed using acridine orange/ethidium bromide staining and enzyme-linked immunosorbent assay-glucose stimulation testing 4 days after cultivation. The results showed that our islet isolation and purification method required 12 ± 3 min, which was significantly shorter than the time required in Ficoll-400, 1077, and HPU groups (34 ± 3, 41 ± 4, and 30 ± 4 min, respectively; P < 0.05). There was no significant difference in islet viability among the four groups. The islet purity, function, yield, and cost of our method were superior to those of the Ficoll-400 and 1077 methods, but inferior to the handpicking method. However, the handpicking method may cause wrist injury and visual impairment in researchers during large-scale islet isolation (>1000 islets). In summary, the MCT method is a rapid, efficient, and economic method for isolating and purifying murine islet cell clumps. This method overcomes some of the shortcomings of conventional methods, showing a relatively higher quality and yield of islets within a shorter duration at a lower cost. Therefore, the current method provides researchers with an alternative option for islet isolation and should be widely generalized.
Highlights
Diabetes is a chronic metabolic disease that affected 400 million patients worldwide in 2014, with an adult prevalence rate of 8.5%, and has become a major threat to human health [1]
We found that the duration of the multi-layer centrifuge tube (MCT) group was 12 ± 3 min, while the durations of the Ficoll-400 group, 1077 group, and hand picking up (HPU) group were 34 ± 3, 41 ± 4, and 30 ± 4 min, respectively
The results showed that in the MTC and HPU groups, very few cells were stained blue with islet viability greater than 98%, while islet viabilities in the Ficoll-400 and 1077 groups were 96.5% and 96.9%, respectively
Summary
Diabetes is a chronic metabolic disease that affected 400 million patients worldwide in 2014, with an adult prevalence rate of 8.5%, and has become a major threat to human health [1]. Efficient, and economic method for isolating and purifying islets from donors remains an important limitation of studies related to islet transplantation and the molecular pathology of diabetes [2]. To overcome this issue, researchers have attempted to improve the isolation and purification method in order to establish a standard operating procedure [3, 4]. We introduce our improved patented device and method for islet isolation and purification using mice as an experimental animal model [6]. We compared the advantages and disadvantages of our method with several conventional methods to achieve efficient, economic, and rapid isolation and purification of islets for use in research studies
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