目的:分离的胰岛是广泛研究的宝贵资源,包括细胞替代研究和基于细胞的平台,用于糖尿病药物发现和疾病建模。胰岛分离是一个复杂而逐步的过程,旨在获得纯净的,可行的,和功能性胰岛的体外和体内研究。应该注意的是,啮齿动物品系的差异,性别,体重,和密度梯度可能会影响孤立的胰岛的属性。我们评估了影响大鼠胰岛分离程序的变量,以达到最大胰岛产量和功能,这对胰岛再生生物学的进一步研究至关重要。
方法:本实验研究比较了两种不同品系的非糖尿病大鼠胰岛的产量和纯度。接下来,比较了男性和女性之间的胰岛颗粒数(IPN)和胰岛当量(IEQ),并研究了产生胰岛数量最多的重量范围。此外,三种不同密度梯度的影响,即Histopaque,Pancoll,和Lymphodex,对最终分离的胰岛纯度和产率进行评估。最后,测量分离的胰岛的活力和功能。
结果:IEQ,IPN,15只李斯特带帽大鼠(LHRs)的分离胰岛纯度显着(P≤0.05)高于其他菌株。男性LHR导致IEQ显著高于女性(P≤0.05)。此外,IPN和IEQ在不同体重组之间没有显着差异。此外,利用Histopaque和Pancoll导致更高的产率和纯度。分离的胰岛的体内评估在移植后第2-5天显示移植组中血糖百分比显著降低。
结论:基于这些结果,以恒定产量分离高质量大鼠胰岛的最佳方案,纯度,和功能已被确立为发展糖尿病研究的重要平台。
OBJECTIVE: Isolated pancreatic islets are valuable resources for a wide range of research, including cell replacement studies and cell-based platforms for diabetes drug discovery and disease modeling. Islet isolation is a complex and stepwise procedure aiming to obtain pure, viable, and functional islets for in vitro and in vivo studies. It should be noted that differences in rodent strains, gender, weight, and density gradients may affect the isolated islet\'s properties. We evaluated the variables affecting the rat islet isolation procedure to reach the maximum islet yield and functionality, which would be critical for further studies on islet regenerative biology.
METHODS: The present experimental
study compared the yield and purity of isolated islets from nondiabetic rats of two different strains. Next, islet particle number (IPN) and islet equivalent (IEQ) were compared between males and females, and the weight range that yields the highest number of islets was investigated. Moreover, the influence of three different density gradients, namely Histopaque, Pancoll, and Lymphodex, on final isolated islets purity and yield were assessed. Finally, the viability and functionality of isolated islets were measured.
RESULTS: The IEQ, IPN, and purity of isolated islets in 15 Lister hooded rats (LHRs) were significantly (P≤0.05) higher than those of the other strains. Male LHRs resulted in significantly higher IEQ compared to females (P≤0.05). Moreover, IPN and IEQ did not significantly vary among different weight groups. Also, the utilization of Histopaque and Pancoll leads to higher yield and purity. In vivo assessments of the isolated islets presented significantly reduced blood glucose percentage in the transplanted group on days 2-5 following transplantation.
CONCLUSIONS: Based on these results, an optimal protocol for isolating high-quality rat islets with a constant yield, purity, and function has been established as an essential platform for developing diabetes research.