PDF(Pubmed)
Abstract：
Pancreatic islet isolation is critical for type 2 diabetes research. Although -omics approaches have shed light on islet molecular profiles, inconsistencies persist; on the other hand, functional studies are essential, but they require reliable and standardized isolation methods. Here, we propose a simplified protocol applied to very small-sized samples collected from partially pancreatectomized living donors. Islet isolation was performed by digesting tissue specimens collected during surgery within a collagenase P solution, followed by a Lympholyte density gradient separation; finally, functional assays and staining with dithizone were carried out. Isolated pancreatic islets exhibited functional responses to glucose and arginine stimulation mirroring donors\' metabolic profiles, with insulin secretion significantly decreasing in diabetic islets compared to non-diabetic islets; conversely, proinsulin secretion showed an increasing trend from non-diabetic to diabetic islets. This novel islet isolation method from living patients undergoing partial pancreatectomy offers a valuable opportunity for targeted study of islet physiology, with the primary advantage of being time-effective and successfully preserving islet viability and functionality. It enables the generation of islet preparations that closely reflect donors\' clinical profiles, simplifying the isolation process and eliminating the need for a Ricordi chamber. Thus, this method holds promises for advancing our understanding of diabetes and for new personalized pharmacological approaches.
摘要：
胰岛隔离是2型糖尿病研究的关键。尽管组学方法已经揭示了胰岛分子谱，矛盾仍然存在；另一方面，功能研究是必不可少的，但是它们需要可靠和标准化的隔离方法。这里,我们提出了一种简化方案,适用于从部分胰腺切除的活体供体采集的非常小的样本.通过在胶原酶P溶液中消化手术期间收集的组织标本来进行胰岛分离。然后进行淋巴细胞密度梯度分离；最后，进行了功能测定和双硫zone染色。分离的胰岛表现出对葡萄糖和精氨酸刺激的功能反应，反映了供体的代谢谱，与非糖尿病胰岛相比，糖尿病胰岛的胰岛素分泌显着减少；相反，胰岛素原分泌从非糖尿病胰岛到糖尿病胰岛呈增加趋势。这种从接受部分胰腺切除术的活体患者中分离胰岛的新方法为有针对性地研究胰岛生理学提供了宝贵的机会。主要优点是时间有效并成功保持胰岛活力和功能。它能够产生密切反映供体临床概况的胰岛制剂，简化了隔离过程，消除了对Ricordi室的需求。因此，这种方法有望促进我们对糖尿病的理解和新的个性化药理学方法。
