成人和脐带血NK细胞的纯化、培养和CD19-CAR慢病毒转导

Q2 Immunology and Microbiology
Lara Herrera, Manel Juan, Cristina Eguizabal
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引用次数: 8

摘要

自然杀伤细胞,或NK细胞,是一种对先天免疫系统至关重要的细胞毒性淋巴细胞。NK细胞的作用类似于细胞毒性T细胞,它们对病毒感染的细胞和肿瘤形成提供快速反应。长期以来,未经修饰的NK细胞一直用于各种免疫疗法来治疗不同的肿瘤,但收效甚微。然而,在过去的几年里,NK细胞修饰表达嵌合抗原受体(CAR-NK细胞)已经成为抗原特异性抗肿瘤药物的特别理想的细胞平台。与CAR-T细胞不同,它们不会引起同种异体反应或移植物抗宿主病,因此可以用于不同MHC表达的受体。本文概述了获得CD19-CAR-NK细胞的方案,重点介绍了获得和培养纯化NK细胞群的重要性,以及如何获得良好的转染效率。©2020 Wiley期刊公司基本方案1:成人外周血和脐带血NK细胞的纯化和培养基本方案2:成人外周血或脐带血NK细胞的CD19-CAR慢病毒转导支持方案:慢病毒上清的生产
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Purification, Culture, and CD19-CAR Lentiviral Transduction of Adult and Umbilical Cord Blood NK Cells

Natural killer cells, or NK cells, are a type of cytotoxic lymphocyte critical to the innate immune system. The role that NK cells play is analogous to that of cytotoxic T cells in that they provide rapid responses to virus-infected cells and responses to tumor formation. Unmodified NK cells have long been used in various immunotherapies to treat different tumors, with only marginal success. However, in the last few years, NK cells modified to express chimeric antigen receptors (CAR-NK cells) have emerged as particularly ideal cellular platforms for antigen-specific antitumor agents. Unlike CAR-T cells, they do not elicit allogeneic responses or graft-versus-host disease and therefore can be administered to recipients with differing MHC expression. This article outlines protocols to obtain CD19-CAR-NK cells, focusing on the importance of obtaining and culturing a purified NK cell population and how to attain good transfection efficiency. © 2020 Wiley Periodicals LLC

Basic Protocol 1: Purification and culture of adult peripheral blood and umbilical cord blood NK cells

Basic Protocol 2: CD19-CAR lentiviral transduction of adult peripheral blood or umbilical cord blood NK cells

Support Protocol: Production of lentiviral supernatant

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来源期刊
Current Protocols in Immunology
Current Protocols in Immunology Immunology and Microbiology-Immunology
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