Application of “Tissueoid Cell Culture System” Using a Silicate Fiber Scaffold for Cancer Research

Abstract

Background: We developed a 3-dimensional (3D) culture system using a high-purity silica fiber scaffold of unwoven sheets called Cellbed TM. Methods: 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. Results: 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. Conclusion: 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.

Differences in the Central Energy Metabolism of Cancer Cells between Conventional 2D and Novel 3D Culture Systems

“…Therefore, it remains challenging to perform the 3D culture of fibroblasts and vascular endothelial cells in Cellbed. Nonetheless, it is to some extent possible to simulate the interstitium by coating Cellbed with collagen, laminin and fibronectin [ 10 ]. The function of 3D-cultured rat hepatocytes grown in Cellbed and coated with these lasts longer in 3D than in 2D [ 35 ].…”

“…Although there are various types of scaffold materials available, we recently developed a tissueoid cell culture system [ 10 ] using a 3D culture scaffold Cellbed TM (Japan Vilene, Tokyo, Japan) that consists of highly pure silicate fiber. Previous 3D culture studies of various cancer cell lines, such as colorectal cancer, Barrett′s adenocarcinoma, gastroesophageal junction cancer, gastric cancer and tongue cancer, have shown that 3D culture systems allow good morphological observations resembling in vivo conditions [ 10 ]. Cellbed is an approximately 200-µm-thick ultrafine nonwoven fiber scaffold [ 11 ].…”

“…Cancer cells change their morphology and move through the pores with a mean diameter of 7–8 µm, which is measured by the bubble point and a mean flow pore test. Compared to the conventional 3D culture systems, the 3D culture system of Cellbed has the following advantages: (I) the absence of other biogenic components, which allows the exclusive study of cancer cells; (II) ease of studying cancer cells similar to 2D culture systems; (III) long-term culture capability; (IV) chemical stability; and (V) 3D culture mounted with a mounting medium with a refraction index equivalent to that of a silicate fiber, which leads to the transparency of Cellbed, allowing postculture observation of the cells with a light microscope [ 10 , 11 ].…”

Similarities and differences in metabolites of tongue cancer cells among two‐ and three‐dimensional cultures and xenografts

“…Recently, there have been reports on 3D cancer organoids and cancer tissue‐originated spheroids using ECM matter such as spheroids and collagen 6 . We also have recently proposed a novel 3D “tissueoid cell culture system” using Cellbed (Japan Vilene Co.) as a 3D culture carrier 7,8 . Cellbed is a fiber aggregate made of ultrafine silica fiber, and its structure is similar to that of the tissue skeleton of loose connective tissue in a living body 7,8 .…”

Cancer Stem Cell Microenvironment Models with Biomaterial Scaffolds In Vitro

“…Silica fibers coated with different types of collagen were used to culture different types of cancer cells in the 3D environment where cells showed enhancement of invasiveness and proliferation when the fibers were coated with collagen IV. This method was called "tissueoid cell culture system" and suggested for its advantages of combability with different cell staining methods [98]. Cell culture vessels were also modified into hydrophobic fluoro-silica (FS) surface resulting in low adherence and transient aggregation of breast cancer cells followed by disaggregation.…”