Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Department of Internal Medicine, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Pancreatology. 2018 Dec;18(8):913-927. doi: 10.1016/j.pan.2018.09.009. Epub 2018 Sep 26.
/Objective: The conventional models currently used to evaluate various anti-tumor therapeutic agents are not sufficient for representing human pancreatic ductal adenocarcinoma (PDA), which has a unique tumor microenvironment. We aimed to produce an organotypic slice culture model from human PDA that resembles the in vivo situation and to evaluate the responses of PDA slices to established cytotoxic drugs.
PDA tissues were obtained from 10 patients who underwent pancreatic resection. The tissues were sliced by a vibratome, and the tumor slices were then cultured. The viability of tumor slices during slice culture was evaluated using H&E and immunohistochemical staining, and stromal cells were demonstrated. The effects of cytotoxic drugs on PDA cell lines and slices were analyzed.
Tumor slices maintained their surface areas and tissue viability for at least five days during culture. Preserved proliferation and apoptosis in tumor slices were observed by the expression of Ki-67 and cleaved caspase-3. Stromal cells including macrophages (CD68 and CD163), T cells (CD3, CD8, and FOXP3), and myeloid cells (CD11b) were present throughout the culture period. Staurosporine, gemcitabine, and cisplatin treatment of PDA cell lines and tumor slices exerted proportional cytotoxic effects in terms of MTT viability, tumor cell number, and Ki-67 and cleaved caspase-3 expression.
Organotypic human PDA slice cultures preserved their viability and tumor microenvironment for at least five days during slice culture. PDA slice culture appears to be a feasible preclinical test model to assess the response to anti-tumor agents.
目的：目前用于评估各种抗肿瘤治疗药物的常规模型不足以代表具有独特肿瘤微环境的人胰腺导管腺癌 (PDA)。我们旨在从人 PDA 中产生类似于体内情况的器官型切片培养模型，并评估 PDA 切片对已建立的细胞毒性药物的反应。
从 10 名接受胰腺切除术的患者中获得 PDA 组织。组织通过振动切片机切片，然后培养肿瘤切片。使用 H&E 和免疫组织化学染色评估切片培养过程中肿瘤切片的活力，并显示基质细胞。分析细胞毒性药物对 PDA 细胞系和切片的影响。
肿瘤切片在培养期间至少保持其表面积和组织活力五天。Ki-67 和 cleaved caspase-3 的表达表明肿瘤切片中观察到增殖和凋亡的保留。在整个培养期间，存在包括巨噬细胞 (CD68 和 CD163)、T 细胞 (CD3、CD8 和 FOXP3) 和髓样细胞 (CD11b) 在内的基质细胞。用 staurosporine、gemcitabine 和 cisplatin 处理 PDA 细胞系和肿瘤切片，在 MTT 活力、肿瘤细胞数量以及 Ki-67 和 cleaved caspase-3 表达方面均表现出比例细胞毒性作用。
器官型人 PDA 切片培养物在切片培养期间至少保持其活力和肿瘤微环境五天。PDA 切片培养似乎是评估抗肿瘤药物反应的可行临床前测试模型。
