Pigni Matteo, Ashok Devika, Acha-Orbea Hans
Department of Biochemistry CIIL, University of Lausanne, Chemin des Boveresses 155, CH-1066, Epalinges, Switzerland.
Methods Mol Biol. 2016;1423:39-49. doi: 10.1007/978-1-4939-3606-9_3.
It is notoriously difficult to obtain large quantities of non-activated dendritic cells ex vivo. For this reason, we produced and characterized a mouse model expressing the large T oncogene under the CD11c promoter (Mushi mice), in which CD8α(+) dendritic cells transform after 4 months. We derived a variety of stable cell lines from these primary lines. These cell lines reproducibly share with freshly isolated dendritic cells most surface markers, mRNA and protein expression, and all tested biological functions. Cell lines can be derived from various strains and knockout mice and can be easily transduced with lentiviruses. In this article, we describe the derivation, culture, and lentiviral transduction of these dendritic cell lines.
在体外获得大量未活化的树突状细胞非常困难，这是出了名的。因此，我们构建并鉴定了一种在CD11c启动子下表达大T癌基因的小鼠模型（Mushi小鼠），其中CD8α(+)树突状细胞在4个月后会发生转化。我们从这些原代细胞系中获得了多种稳定的细胞系。这些细胞系与新鲜分离的树突状细胞可重复性地共享大多数表面标志物、mRNA和蛋白质表达以及所有测试的生物学功能。细胞系可源自各种品系和基因敲除小鼠，并且可以很容易地用慢病毒进行转导。在本文中，我们描述了这些树突状细胞系的衍生、培养和慢病毒转导。
