Division of Molecular and Diagnostic Pathology, Department of Pathology, Shiga University of Medical Science, Otsu, Japan.
Department of Oral and Maxillofacial Surgery, Shiga University of Medical Science, Otsu, Japan.
Pathobiology. 2020;87(5):291-301. doi: 10.1159/000509133. Epub 2020 Sep 23.
We developed a 3-dimensional (3D) culture system using a high-purity silica fiber scaffold of unwoven sheets called CellbedTM.
We used adherent colon and esophagogastric junction adenocarcinoma cells, tongue squamous cell carcinoma (SqCC) cells, and nonadherent gastric cancer cells. These cells were subjected to staining with various substances and observed by electron microscopy. To evaluate the effects of extracellular matrix in carcinoma tissues, SqCC cells were cultured in Cellbed coated with collagens I, III, and IV.
Especially well-differentiated carcinoma cells cultured in this 3D system showed their own unique characteristics: luminal formation in adenocarcinoma cells and cell stratification and keratinization in SqCC cells. Scanning electron microscopy revealed the proliferation of cancer cells with cytoplasm entwined in Cellbed. Intercellular desmosomes in squamous epithelia were detected by transmission electron microscopy of vertical cross sections. SqCC cells cultured in Cellbed coated with collagen IV showed enhanced invasive and proliferative abilities.
Because the morphology of cancer cells cultured in this 3D culture system is similar to that in living organisms, we called the system a "tissueoid cell culture system." Coating with collagen IV enables the modification of cell-matrix interactions as well as recapitulation of the in vivo microenvironment.
我们开发了一种使用称为 CellbedTM 的无纺织物片高纯度二氧化硅纤维支架的 3 维(3D)培养系统。
我们使用贴壁结肠和食管胃交界腺癌细胞、舌鳞癌细胞(SqCC)和非贴壁胃癌细胞。这些细胞用各种物质进行染色,并通过电子显微镜观察。为了评估细胞外基质在癌组织中的作用,将 SqCC 细胞培养在涂有胶原蛋白 I、III 和 IV 的 Cellbed 上。
特别是在这个 3D 系统中培养的分化良好的癌细胞表现出其自身独特的特征:腺癌细胞中的腔形成和 SqCC 细胞中的细胞分层和角化。扫描电子显微镜显示出癌细胞在 Cellbed 中增殖,细胞质交织在一起。通过垂直横截面的透射电子显微镜检测到鳞状上皮细胞的细胞间桥粒。在涂有胶原蛋白 IV 的 Cellbed 上培养的 SqCC 细胞显示出增强的侵袭和增殖能力。
由于在这个 3D 培养系统中培养的癌细胞的形态与生物体内的形态相似,我们称该系统为“组织样细胞培养系统”。胶原蛋白 IV 的涂层能够修饰细胞-基质相互作用,并重现体内微环境。