Shupe Thomas D, Piscaglia Anna C, Oh Seh-Hoon, Gasbarrini Antonio, Petersen Bryon E
Department of Pathology, Immunology and Laboratory Medicine, College of Medicine, University of Florida, Gainesville, FL, USA.
Methods Mol Biol. 2009;482:387-405. doi: 10.1007/978-1-59745-060-7_24.
The pace of research on the potential therapeutic uses of liver stem cells or "oval cells" has accelerated significantly in recent years. Concurrent advancements in techniques for the isolation and characterization of these cells have helped fuel this research. Several models now exist for the induction of oval cell proliferation in rodents. Protocols for the isolation and culture of these cells have evolved to the point that they may be set up in any laboratory equipped for cell culture. The advent of magnetic cell sorting has eliminated reliance on expensive flow cytometric sorting equipment to generate highly enriched populations of oval cells. Our laboratory has had much success in using the oval cell surface marker Thy-1 in combination with magnetic sorting to produce material suitable for testing the influence of a myriad of chemical signaling molecules on the oval cell phenotype. This chapter will describe our basic strategy for oval cell induction and isolation. Additionally, two in vitro procedures are described which the reader may find useful in the early stages of developing an oval cell research project.
近年来,关于肝干细胞或“卵圆细胞”潜在治疗用途的研究步伐显著加快。这些细胞分离和鉴定技术的同步进步推动了这项研究。目前有几种在啮齿动物中诱导卵圆细胞增殖的模型。这些细胞的分离和培养方案已经发展到在任何具备细胞培养条件的实验室都可以建立的程度。磁性细胞分选技术的出现消除了对昂贵的流式细胞分选设备的依赖,从而能够获得高度富集的卵圆细胞群体。我们实验室在使用卵圆细胞表面标志物Thy-1结合磁性分选来制备适合测试多种化学信号分子对卵圆细胞表型影响的材料方面取得了很大成功。本章将描述我们诱导和分离卵圆细胞的基本策略。此外,还描述了两种体外实验方法,读者可能会发现它们在开展卵圆细胞研究项目的早期阶段很有用。