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使用组织样细胞培养系统进行胶质母细胞瘤细胞的三维培养

Three-Dimensional Culture of Glioblastoma Cells Using a Tissueoid Cell Culture System.

作者信息

Okamoto Natsume, Taniura Naoko, Nakayama Takahisa, Tanaka Eri, Kageyama Yusuke, Noujima Mai, Kushima Ryoji, Mukaisho Ken-Ichi

机构信息

Division of Human Pathology, Department of Pathology, Shiga University of Medical Science, Otsu, Japan.

Education Center for Medicine and Nursing, Shiga University of Medical Science, Otsu, Japan.

出版信息

Acta Histochem Cytochem. 2024 Oct 28;57(5):149-155. doi: 10.1267/ahc.24-00043. Epub 2024 Oct 12.

DOI:10.1267/ahc.24-00043
PMID:39552931
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11565224/
Abstract

In classical cell culture techniques, cancer cells typically proliferate in a single layer by adhering to the undersurface of laboratory vessels. Consequently, concerns have been raised regarding the fidelity of the morphological and functional characteristics of these cultured cancer cells compared to those of their counterparts. Our previous studies have investigated various epithelial malignant tumors utilizing the Tissueoid cell culture system, a three-dimensional (3D) cultivation method employing Cellbed-a nonwoven sheet composed of high-purity silica fibers as a scaffold. In this investigation, we have achieved successful 3D culturing of glioblastoma cells (A172 and T98G), which are non-epithelial in nature. As such our focus is to juxtapose their morphological features against that of those cultivated via conventional two-dimensional (2D) methods. Our findings will be elucidated using immunostaining, immunofluorescence staining, and scanning electron microscopy, substantiated with accompanying imaging. Notably, cells cultured in the 3D environment exhibited distinct morphological attributes compared to those of their 2D counterparts, notably featuring pronounced cellular protrusions. We envisage the continued utilization of the 3D culture platform to facilitate diverse avenues of research, encompassing the exploration of novel therapeutic modalities for glioblastoma cells and beyond.

摘要

在经典的细胞培养技术中,癌细胞通常通过附着在实验室容器的底面以单层形式增殖。因此,与对应肿瘤细胞相比,这些培养的癌细胞的形态和功能特征的保真度受到了关注。我们之前的研究利用组织样细胞培养系统对各种上皮恶性肿瘤进行了研究,该系统是一种三维(3D)培养方法,采用Cellbed(一种由高纯度二氧化硅纤维组成的无纺布作为支架)。在本研究中,我们成功地对本质上为非上皮性的胶质母细胞瘤细胞(A172和T98G)进行了3D培养。因此,我们的重点是将它们的形态特征与通过传统二维(2D)方法培养的细胞的形态特征进行对比。我们的研究结果将通过免疫染色、免疫荧光染色和扫描电子显微镜进行阐释,并辅以相关成像。值得注意的是,与二维培养的细胞相比,在三维环境中培养的细胞表现出明显不同的形态特征,特别是具有明显的细胞突起。我们设想继续利用三维培养平台来促进多样化的研究途径,包括探索针对胶质母细胞瘤细胞及其他细胞的新型治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/1152ed719c87/AHC24-00043f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/169c222de4d3/AHC24-00043f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/8ff21df2bfeb/AHC24-00043f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/9069d9c88d9f/AHC24-00043f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/cab144350c84/AHC24-00043f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/0d51e01dba7c/AHC24-00043f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/1152ed719c87/AHC24-00043f06.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/169c222de4d3/AHC24-00043f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/8ff21df2bfeb/AHC24-00043f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/9069d9c88d9f/AHC24-00043f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/cab144350c84/AHC24-00043f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/0d51e01dba7c/AHC24-00043f05.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/71f9/11565224/1152ed719c87/AHC24-00043f06.jpg

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本文引用的文献

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Cancer Cell. 2024 Jul 8;42(7):1217-1238.e19. doi: 10.1016/j.ccell.2024.06.004.
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Silicate Microfiber Scaffolds Support the Formation and Expansion of the Cortical Neuronal Layer of Cerebral Organoids With a Sheet-Like Configuration.硅酸钙微纤维支架支持具有片状结构的脑类器官皮质神经元层的形成和扩展。
Stem Cells Transl Med. 2023 Dec 18;12(12):825-837. doi: 10.1093/stcltm/szad066.
3
Towards a New 3Rs Era in the construction of 3D cell culture models simulating tumor microenvironment.
迈向模拟肿瘤微环境的3D细胞培养模型构建中的新3R时代。
Front Oncol. 2023 Apr 3;13:1146477. doi: 10.3389/fonc.2023.1146477. eCollection 2023.
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Recreating the extracellular matrix: novel 3D cell culture platforms in cancer research.重建细胞外基质:癌症研究中的新型 3D 细胞培养平台。
FEBS J. 2023 Nov;290(22):5238-5247. doi: 10.1111/febs.16778. Epub 2023 Mar 27.
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Glioblastoma and Other Primary Brain Malignancies in Adults: A Review.成人脑胶质瘤和其他原发性脑恶性肿瘤:综述。
JAMA. 2023 Feb 21;329(7):574-587. doi: 10.1001/jama.2023.0023.
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Morphologic Analysis of M2 Macrophage in Glioblastoma: Involvement of Macrophage Extracellular Traps (METs).胶质母细胞瘤中M2巨噬细胞的形态学分析:巨噬细胞胞外陷阱(METs)的参与
Acta Histochem Cytochem. 2022 Aug 27;55(4):111-118. doi: 10.1267/ahc.22-00018. Epub 2022 Aug 10.
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3D modeling in cancer studies.癌症研究中的三维建模。
Hum Cell. 2022 Jan;35(1):23-36. doi: 10.1007/s13577-021-00642-9. Epub 2021 Nov 10.
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Advancement of Scaffold-Based 3D Cellular Models in Cancer Tissue Engineering: An Update.基于支架的3D细胞模型在癌症组织工程中的进展:最新情况
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