Protection against apoptosis of islet cells:a combination of Caspase and Cocktail protease inhibitors

Tian Ting-ting; Zhao Shan-shan; Wang Qun; Ye Bin

doi:10.3969/j.issn.2095-4344.2015.37.012

Abstract：BACKGROUND:Currently, there are few reports on the effects of Caspase inhibitors combined with Cocktail protease inhibitors on isolation and purification of islet cel s. OBJECTIVE:To explore the effects of Caspase inhibitors and Cocktail protease as protectors on islet cel s during isolation and purification process. METHODS:New-born pigs were selected to separate, purify and culture the islet cel s, and after 24 hours of culture, the specimens were divided into three groups:blank control group, experimental group A (Caspase inhibitors and Cocktail protease were added only during digestion process), and experimental group B (Caspase inhibitors and Cocktail protease were added both during digestion and culture processes). AO-EB staining was used to qualitatively observe cel morphology and apoptosis. Flow cytometry was employed to quantitatively detect cel viability and apoptosis. RESULTS AND CONCLUSION:Percentages ofβcel s were 66.91%in blank control group, 84.58%in the experimental group A, and 87.15%in the experimental group B. The proportions of living cel s, apoptotic cel s and dead cel s were 56.52%, 16.15%, 21.25%in the blank control group, 62.27%, 14.66%, 14.47%in the experimental group A, and 73.09%, 6.83%, 10.28%in the experimental group B, respectively. These findings indicate that in the process of digestion and culture in vitro, Caspase inhibitors and Cocktail protease inhibitors can both obviously decrease the loss of cel s, and increase the percentage of islet beta cel s.