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为进行钙成像、细胞损伤测量和腺病毒感染而制备胰腺腺泡细胞。

Preparation of pancreatic acinar cells for the purpose of calcium imaging, cell injury measurements, and adenoviral infection.

作者信息

Orabi Abrahim I, Muili Kamaldeen A, Wang Dong, Jin Shunqian, Perides George, Husain Sohail Z

机构信息

Rangos Research Center, Pediatric Gastroenterology, Hepatology, and Nutrition, Children's Hospital of Pittsburgh of UPMC.

出版信息

J Vis Exp. 2013 Jul 5(77):e50391. doi: 10.3791/50391.

Abstract

The pancreatic acinar cell is the main parenchymal cell of the exocrine pancreas and plays a primary role in the secretion of pancreatic enzymes into the pancreatic duct. It is also the site for the initiation of pancreatitis. Here we describe how acinar cells are isolated from whole pancreas tissue and intracellular calcium signals are measured. In addition, we describe the techniques of transfecting these cells with adenoviral constructs, and subsequently measuring the leakage of lactate dehydrogenase, a marker of cell injury, during conditions that induce acinar cell injury in vitro. These techniques provide a powerful tool to characterize acinar cell physiology and pathology.

摘要

胰腺腺泡细胞是外分泌胰腺的主要实质细胞,在将胰腺酶分泌到胰管中起主要作用。它也是胰腺炎起始的部位。在此我们描述从整个胰腺组织中分离腺泡细胞并测量细胞内钙信号的方法。此外,我们描述用腺病毒构建体转染这些细胞的技术,以及随后在体外诱导腺泡细胞损伤的条件下测量乳酸脱氢酶(一种细胞损伤标志物)泄漏的技术。这些技术为表征腺泡细胞的生理学和病理学提供了强大的工具。

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