A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery

Sabine Hensler, Claudia Kuehlbach, B. Kotkamp, Dario Frey, M. Mueller
{"title":"A Novel Standardized Inflammatory Cell-Modulated 3D Tumor Tissue Model for Analysis of Tumor-Stroma Interaction and Drug Discovery","authors":"Sabine Hensler, Claudia Kuehlbach, B. Kotkamp, Dario Frey, M. Mueller","doi":"10.11648/J.BIO.20210904.13","DOIUrl":null,"url":null,"abstract":"The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.","PeriodicalId":284331,"journal":{"name":"American Journal of Bioscience and Bioengineering","volume":"57 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2021-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"American Journal of Bioscience and Bioengineering","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.11648/J.BIO.20210904.13","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1

Abstract

The last decades were marked by substantial progress in understanding the role of tumor-supporting inflammatory reactions in tumor growth and progression. While in vivo data substantiate the contribution of the inflammatory infiltrate and of tumor associated fibroblasts in promoting tumor growth and progression, little is known about the dynamic interaction of these two stromal cell types and their reciprocal influence on each other and on the tumor cells. Mechanistical analyses of these crucial interactions require a standardized and easy to manipulate environment. We therefore established a 3D organotypic in vitro model for epithelial tumors to analyze the interaction of macrophages, neutrophils and fibroblasts in the tumor microenvironment of malignant tumors. In the 3D model, epithelial tumor cells are grown on a collagen type I gel containing fibroblasts, macrophages and neutrophils. Comparable to the in vivo setting, the cytokine driven interaction between macrophages and fibroblasts markedly influences invasion and enhances M2 differentiation in the presence of tumor cells. Addition of neutrophils further leads to a strikingly enhanced tumor invasion associated with an increased expression of MMP-9 and a N2 differentiation of neutrophils. Thus, this novel 3D model provides an in vivo like tissue context to analyze tumor stroma interactions and presents an excellent tool for targeted interference. As such, the model is highly suitable for pharmaceutical screening of novel therapeutics. However, the use of collagen type 1 with its known batch to batch variability as ECM equivalent prohibits the model-standardization that is needed for pharmaceutical testing. Therefore, the 3D in vitro tumor-stroma model was adapted to the use of a bioinert dextran-hydrogel providing a highly standardized and easily modifiable scaffold material that allows the recovery of cells after pharmaceutical experiments. Comparable to the collagen-based model, cells maintained their physiological proliferation, migration and differentiation. Utilizing this standardized model, the efficacy and the tissue impact of novel pharmaceuticals can be investigated in detail with respect to cell morphology, behavior, viability as well as gene expression profiles thereby providing a 3D hydrogel tumor stroma a model that is of great interest for the pharmaceutical industry.
一种新的标准化炎症细胞调节的三维肿瘤组织模型,用于分析肿瘤-基质相互作用和药物发现
在过去的几十年里,在了解肿瘤支持炎症反应在肿瘤生长和进展中的作用方面取得了实质性进展。虽然体内数据证实了炎症浸润和肿瘤相关成纤维细胞在促进肿瘤生长和进展中的作用,但对这两种基质细胞类型的动态相互作用以及它们相互作用和对肿瘤细胞的相互影响知之甚少。对这些关键相互作用的机械分析需要标准化和易于操作的环境。因此,我们建立了上皮性肿瘤的三维体外器官型模型,分析巨噬细胞、中性粒细胞和成纤维细胞在恶性肿瘤肿瘤微环境中的相互作用。在3D模型中,上皮肿瘤细胞生长在含有成纤维细胞、巨噬细胞和中性粒细胞的I型胶原凝胶上。与体内环境相比,在肿瘤细胞存在的情况下,巨噬细胞和成纤维细胞之间的细胞因子驱动的相互作用显著影响侵袭并增强M2分化。中性粒细胞的增加进一步导致与MMP-9表达增加和中性粒细胞N2分化相关的显著增强的肿瘤侵袭。因此,这种新颖的3D模型提供了一个活体组织环境来分析肿瘤间质相互作用,并提供了一个很好的靶向干扰工具。因此,该模型非常适合用于新疗法的药物筛选。然而,使用具有已知批次间可变性的1型胶原蛋白作为ECM等效物,禁止了药物测试所需的模型标准化。因此,体外3D肿瘤基质模型适用于使用生物惰性右旋糖酐水凝胶,提供高度标准化且易于修改的支架材料,允许药物实验后细胞的恢复。与基于胶原的模型相比,细胞保持了生理增殖、迁移和分化。利用这个标准化模型,可以从细胞形态、行为、活力以及基因表达谱等方面详细研究新型药物的功效和对组织的影响,从而提供一个3D水凝胶肿瘤基质模型,这是制药行业非常感兴趣的模型。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信