Jiang Xiuyun, Seo Y David, Sullivan Kevin M, Pillarisetty Venu G
Department of Surgery, University of Washington School of Medicine, Seattle, WA, USA.
Methods Mol Biol. 2019;1884:283-295. doi: 10.1007/978-1-4939-8885-3_20.
Although immunotherapy is currently being widely applied to treat a variety of cancers, there is great heterogeneity in the response to these treatments. Many in the field hypothesize that this may be attributable to the characteristics of each individual tumor immune microenvironment, in addition to systemic immune factors. Therefore, understanding the immune cell microenvironment in a variety of tumors is critically important. Specifically, the interactions among immune, stromal, and cancer cells, along with other factors in tumors, may hold the key to developing rational personalized combinations of immunotherapeutic drugs. We recently developed an organotypic slice culture technique, which enables precise study of the pancreatic ductal adenocarcinoma (PDA) tumor microenvironment. We used a Vibratome to cut fresh human tumor tissue into 250 μm thick slices, and cultured slices on cell culture inserts with 0.4 μm pore to produce our tumor slice culture (TSC) system. We showed that TSC maintained many elements of the original tumor microenvironment and architecture for approximately one week. Using this slice culture technique for PDA, we demonstrated that immune cells, including T cells and macrophages, cancer cells, and stromal myofibroblasts were present throughout the culture period. TSCs were functionally responsive to drug treatment. Live PDA slices could be stained for multicolor immunofluorescence imaging of each of the primary cellular constituents of the tumor. Finally, autologous CFSE-labeled splenocytes were observed to readily migrate into cocultured tumor slices.
尽管免疫疗法目前正广泛应用于治疗多种癌症，但这些治疗的反应存在很大的异质性。该领域的许多人推测，除了全身免疫因素外，这可能归因于每个肿瘤免疫微环境的特征。因此，了解各种肿瘤中的免疫细胞微环境至关重要。具体而言，免疫细胞、基质细胞和癌细胞之间的相互作用，以及肿瘤中的其他因素，可能是开发合理的个性化免疫治疗药物组合的关键。我们最近开发了一种器官型切片培养技术，能够精确研究胰腺导管腺癌（PDA）肿瘤微环境。我们使用振动切片机将新鲜的人类肿瘤组织切成250μm厚的切片，并在孔径为0.4μm的细胞培养插入物上培养切片，以产生我们的肿瘤切片培养（TSC）系统。我们表明，TSC在大约一周的时间内维持了原始肿瘤微环境和结构的许多要素。使用这种针对PDA的切片培养技术，我们证明了在整个培养期间都存在免疫细胞（包括T细胞和巨噬细胞）、癌细胞和基质肌成纤维细胞。TSC对药物治疗具有功能反应性。活的PDA切片可以进行染色，以便对肿瘤的每种主要细胞成分进行多色免疫荧光成像。最后，观察到自体CFSE标记的脾细胞能够轻易迁移到共培养的肿瘤切片中。
