Department of Chemistry and Biochemistry, Florida State University, 95 Chieftain Way, Tallahassee, FL 32306, USA.
Anal Methods. 2021 Aug 28;13(32):3614-3619. doi: 10.1039/d1ay00703c. Epub 2021 Jul 26.
Glucagon is a 29-amino acid peptide released from α-cells within pancreatic islets of Langerhans to help raise blood glucose levels. While a plethora of methodologies have been developed for quantitative measurement of insulin released from islets, such methods are not well developed for glucagon despite its importance in blood sugar regulation. In this work, a simple yet robust microfluidic device was developed for holding human pancreatic islets and perfuse them with glucose. The perfusate was collected into 2 min fractions and glucagon quantified using a homogeneous time-resolved Förster resonance energy transfer (TR-FRET) sandwich immunoassay. Simulation of fluid flow within the microfluidic device indicated the device produced low amounts of shear stress on islets, and characterization of the flow with standard glucagon solutions revealed response times within 2 fractions (<4 min). Results with human islets from multiple donors demonstrated either a "burst" of glucagon or a "sustained" glucagon release across the entire period of stimulation. The simplicity, yet robustness, of the device and method is expected to appeal to a number of researchers examining pancreatic islet physiology.
胰高血糖素是一种 29 个氨基酸的肽，从胰岛的α细胞中释放出来，有助于提高血糖水平。尽管胰高血糖素在血糖调节中很重要，但已经开发出了大量用于定量测量胰岛释放的胰岛素的方法，但针对胰高血糖素的方法却并不完善。在这项工作中，开发了一种简单而强大的微流控装置，用于容纳人胰岛并以葡萄糖对其进行灌注。将灌流液收集到 2 分钟的分数中，并使用均相时间分辨荧光共振能量转移（TR-FRET）夹心免疫测定法定量测定胰高血糖素。微流控装置内流体流动的模拟表明，该装置对胰岛产生的剪切应力很小，并且用标准胰高血糖素溶液对流动进行的表征表明，反应时间在 2 个分数内（<4 分钟）。来自多个供体的人胰岛的结果表明，在整个刺激期间，胰高血糖素要么“爆发”，要么“持续”释放。该设备和方法的简单性和强大性有望吸引许多研究人员研究胰岛生理学。
