Islet Cell and Regenerative Biology, Joslin Diabetes Center, Department of Medicine, Brigham and Women's Hospital, Harvard Stem Cell Institute, Harvard Medical School, Boston, MA, USA.
Chemical Biology and Therapeutics Science Program, Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Life Sci Alliance. 2021 Jan 29;4(4). doi: 10.26508/lsa.202000840. Print 2021 Apr.
Highly sensitive approaches to target insulin-expressing cells would allow more effective imaging, sorting, and analysis of pancreatic β-cells. Here, we introduce the use of a reaction-based probe, diacetylated Zinpyr1 (DA-ZP1), to image pancreatic β-cells and β-like cells derived from human pluripotent stem cells. We harness the high intracellular zinc concentration of β-cells to induce a fluorescence signal in cells after administration of DA-ZP1. Given its specificity and rapid uptake by cells, we used DA-ZP1 to purify live stem cell-derived β-like cells as confirmed by immunostaining analysis. We tested the ability of DA-ZP1 to image transplanted human islet grafts and endogenous mouse pancreatic islets in vivo after its systemic administration into mice. Thus, DA-ZP1 enables purification of insulin-secreting β-like cells for downstream applications, such as functional studies, gene-expression, and cell-cell interaction analyses and can be used to label engrafted human islets and endogenous mouse islets in vivo.
高灵敏度的靶向胰岛素表达细胞的方法将允许更有效地对胰腺β细胞进行成像、分选和分析。在这里，我们引入了一种基于反应的探针，双乙酰化 Zinpyr1（DA-ZP1），用于对胰腺β细胞和源自人多能干细胞的β样细胞进行成像。我们利用β细胞内的高锌浓度，在给予 DA-ZP1 后诱导细胞内的荧光信号。鉴于其特异性和快速被细胞摄取，我们使用 DA-ZP1 来纯化活的干细胞衍生的β样细胞，这一点通过免疫染色分析得到了证实。我们测试了 DA-ZP1 在将其全身给药到小鼠体内后，对移植的人胰岛移植物和内源性小鼠胰岛进行体内成像的能力。因此，DA-ZP1 能够用于纯化胰岛素分泌β样细胞，用于下游应用，如功能研究、基因表达和细胞-细胞相互作用分析，并且可以用于标记体内移植的人胰岛和内源性小鼠胰岛。
