Duarte-Sanmiguel Silvia, Higuita-Castro Natalia, Gallego-Perez Daniel
Department of Biomedical Engineering, The Ohio State University, Columbus, OH, USA.
Department of Surgery, The Ohio State University, Columbus, OH, USA.
Methods Mol Biol. 2020;2050:79-84. doi: 10.1007/978-1-4939-9740-4_8.
Dendritic cells (DCs) are cells of the immune system that behave as antigen presenters and assist in T cell activation. DCs have recently been used in cell-based immunotherapies for the treatment of different diseases due to the lack of adverse nonspecific immune responses, typically elicited by other approaches. Genetically modified DCs, for example, have been used to stimulate CD4/CD8 antigen presenting immune responses. However, genetic manipulation of primary DCs remains a challenge. Here we describe a protocol for nonviral, benign transfection of primary DCs using nanochannel-based electroporation, and the subsequent collection of genetically modified exosomes for downstream applications.
树突状细胞（DCs）是免疫系统中的细胞，作为抗原呈递细胞，协助T细胞活化。由于缺乏通常由其他方法引发的不良非特异性免疫反应，DCs最近已被用于基于细胞的免疫疗法来治疗不同疾病。例如，基因改造的DCs已被用于刺激CD4/CD8抗原呈递免疫反应。然而，原代DCs的基因操作仍然是一个挑战。在此，我们描述了一种使用基于纳米通道的电穿孔对原代DCs进行非病毒、良性转染的方案，以及随后收集基因改造的外泌体用于下游应用。
