利用单细胞 Western Blot 平台表征人类胰岛细胞

IF 1.7 4区 医学 Q3 GASTROENTEROLOGY & HEPATOLOGY Pancreas Pub Date : 2024-09-12 DOI:10.1097/mpa.0000000000002385
Gal Lenz,Lynn Miao,Ayelet Lenz,Jacob Mares,Janine Quijano,Heather N Zook,Hirotake Komatsu,Pablo Garcia,Kevin Ferreri,Hsun Teresa Ku,Fouad Kandeel
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引用次数: 0

摘要

目的胰岛移植是治疗 1 型糖尿病的有效方法。然而,移植成功与否取决于能否快速评估胰岛,因为胰岛在分离后 2-3 天就会恶化。一种新工具--单细胞 Western 印迹(scWestern)可在一天内得到结果。在本研究中,我们旨在测试单细胞 Western 印迹法是否适合检测胰岛中三种主要内分泌细胞类型--β(胰岛素)、α(胰高血糖素)和δ(体生长抑素)细胞的蛋白质标记物。方法我们对 scWestern 平台上的抗体特异性、信号强度和细胞识别进行了鉴定,然后比较了 scWestern 和综合胰岛分布计划 (IIDP) 免疫组化进行的胰岛细胞组成分析。单个胰岛细胞中胰岛素、胰高血糖素和体生长抑素的蛋白表达水平差异很大,突出表明了细胞类型的异质性。令人惊讶的是,scWestern 发现了双激素细胞(约 1%),在非糖尿病和非肥胖成人胰岛中共同表达胰岛素和体节素或胰岛素和胰高血糖素,共聚焦免疫荧光显微镜证实了这一点。scWestern平台可在临床移植前及时评估离体胰岛中β细胞的数量。
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Characterization of Human Pancreatic Islet Cells using a Single-Cell Western Blot Platform.
OBJECTIVE Islet transplantation is an effective treatment for type 1 diabetes. However, transplant success depends on quick islet assessment because islets deteriorate 2-3 days after isolation. A new tool, single-cell Western blot (scWestern), offers results within one day. In this study, we aimed to test the suitability of scWestern to detect protein markers for beta (insulin), alpha (glucagon), and delta (somatostatin) cells, the three major endocrine cell types in islets. METHODS We characterized the antibody specificity, signal intensity, and cell identification on the scWestern platform, and then compared the islet cell composition analysis between scWestern and immunohistochemistry performed by the Integrated Islet Distribution Program (IIDP). RESULTS Islet cell composition is comparable for alpha and beta cells, but not delta cells. Protein expression levels of insulin, glucagon, and somatostatin in individual islet cells varied greatly, highlighting cell type heterogeneity. Surprisingly, scWestern revealed double-hormonal cells (~1%), co-expressing insulin and somatostatin or insulin and glucagon, in non-diabetic and non-obese adult human islets, which was confirmed by confocal immunofluorescence microscopy. CONCLUSIONS These results demonstrate that each alpha, beta, and delta cells express varying levels of peptide hormones, and a small subpopulation co-expresses double hormones in normal human islets. The scWestern platform will enable timely assessment of beta cell mass in isolated islets before clinical transplantation.
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来源期刊
Pancreas
Pancreas 医学-胃肠肝病学
CiteScore
4.70
自引率
3.40%
发文量
289
审稿时长
1 months
期刊介绍: Pancreas provides a central forum for communication of original works involving both basic and clinical research on the exocrine and endocrine pancreas and their interrelationships and consequences in disease states. This multidisciplinary, international journal covers the whole spectrum of basic sciences, etiology, prevention, pathophysiology, diagnosis, and surgical and medical management of pancreatic diseases, including cancer.
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