Isolation of tumor endothelial cells from murine cancer
Isolation of tumor endothelial cells from murine cancer
复制标题
从小鼠癌症中分离肿瘤内皮细胞
DOI：
10.1016/j.jim.2018.11.005
复制
发表时间：
2019
影响因子：
2.2
通讯作者：
Ohdan Hideki
中科院分区：
文献类型：
--
作者：
Taguchi Kazuhiro;Onoe Takashi;Yoshida Tomoaki;Yamashita Yoshinori;Taniyama Kiyomi;Ohdan Hideki
肿瘤内皮细胞（TEC），构成肿瘤血管的衬里，具有作为肿瘤组成细胞的各种特性。在这项研究中，我们描述了一种分离高纯度新鲜TEC的新方法，这些TEC在肿瘤内形成一个小群体。首先从荷瘤小鼠解剖肿瘤，并用II型胶原酶消化成单细胞悬液;然后通过密度梯度离心分离单细胞。通过使用磁性激活细胞分选的CD 31阳性选择富集TEC，随后通过荧光激活细胞分选纯化TEC。通过流式细胞术验证所获得的细胞的高纯度。在细胞培养时，分离的细胞显示出多边形形状和鹅卵石外观，这是内皮细胞的特征。此外，功能测定显示TEC在体外抑制CD 8 + T细胞的增殖。我们相信，在这项研究中所描述的分离方法将能够进一步阐明的特征TEC。
Tumor endothelial cells (TECs), which constitute the lining of the tumor blood vessels, have various characteristics as tumor constituent cells. In this study, we describe a novel method for the isolation of highly pure, fresh TECs, which form a small population within the tumor. Tumors were first dissected from tumor-bearing mice and digested to a single cell suspension with Collagenase Type II; the single cells were then separated by density gradient centrifugation. TECs were enriched by CD31-positive selection using magnetic activated cell sorting and subsequently purified by fluorescence activated cell sorting. The high purity of the obtained cells was verified by flow cytometry. Upon cell culture, the isolated cells showed a polygonal shape and a cobblestone appearance, which are features of the endothelial cells. Furthermore, a functional assay revealed that the TECs suppressed the proliferation of CD8+ T cells in vitro. We believe that the isolation method described in this study will enable the further elucidation of the characteristics of TECs.