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DOI: 10.3791/51321-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This article presents a protocol for generating a native matrix from fibroblasts to study tumor cell invasion. The methodology allows for the assessment of the impact of different stromal cell types on tumor cell biology.
组织工程化的成纤维细胞衍生的天然基质是一种新兴的工具来生成支持上皮细胞增殖和分化的基质衬底。这里应用此方法,以评估不同的基质细胞类型的肿瘤细胞生物学的影响的协议被提出。
以下程序的总体目标是从成纤维细胞生成天然基质,用于评估肿瘤细胞侵袭。首先通过在富含山梨酸的培养基中在塑料上培养成纤维细胞以生成天然基质来实现。在第二步中,天然基质从塑料生长表面分离并释放,然后将该基质接种肿瘤细胞,并在固定时间后提升到气液界面。
收获这些 3D 培养物用于肿瘤细胞侵袭的组织学分析。最终,可以通过免疫组织学分析来评估天然成纤维细胞基质对肿瘤细胞侵袭的影响。因此,这种技术生成 3D 典型培养物的优势在于,我们可以在没有合成或外来底物的情况下评估成纤维细胞本身产生的天然基质,例如跨物种 1 型胶原蛋白首先将 200, 000 个成纤维细胞接种在成纤维细胞培养基中的六个孔板中,补充有新鲜制备的 IC 酸。
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