Organ-like microenviroment and 3-dimensional (3D) cell culture conformations have been suggested as promising approaches to mimic in a micro-scale a whole organ cellular functions and interactions present in vivo. We have used this approach to examine biologic features of hepatocellular carcinoma (HCC) cells. In this study, we demonstrate that hepatocellular carcinoma (HCC) cells, fibroblasts, endothelial cells and extracellular matrix can generate organoid-like spheroids that enhanced numerous features of human HCC observed in vivo. We show that the addition of non-parenchymal cells such as fibroblast and endothelial cells is required for spheroid formation as well as the maintenance of the tissue-like structure. Furthermore, HCC cells cultured as spheroids with non-parenchymal cells express more neo-angiogenesis-related markers (VEGFR2, VEGF, HIF-α), tumor-related inflammatory factors (CXCR4, CXCL12, TNF-α) and molecules-related to induced epithelial-mesenchymal transition (TGFβ, Vimentin, MMP9) compared with organoids containing only HCC cells.

These results demonstrate the importance of non-parenchymal cells in the cellular composition of HCC organoids. The novelty of the multicellular-based organotypic culture system strongly supports the integration of this approach in a high throughput approach to identified patient-specific HCC malignancy and accurate anti-tumor therapy screening after surgery.

已提出器官样微环境和3维（3D）细胞培养构象是在微观规模上模拟体内存在的整个器官细胞功能和相互作用的有前途的方法。我们已经使用这种方法来检查肝细胞癌（HCC）细胞的生物学特征。在这项研究中，我们证明肝细胞癌（HCC）细胞，成纤维细胞，内皮细胞和细胞外基质可以产生类器官样球体，从而增强体内观察到的人类HCC的众多特征。我们表明，非实质细胞如成纤维细胞和内皮细胞的添加是球体形成以及组织样结构维持所必需的。此外，与非实质细胞培养成球状的HCC细胞表达更多的新血管生成相关标记（VEGFR2，VEGF，

这些结果证明了非实质细胞在HCC类器官的细胞组成中的重要性。基于多细胞的器官培养系统的新颖性强烈支持该方法以高通量方法集成，以识别患者特定的HCC恶性肿瘤并在手术后进行准确的抗肿瘤治疗筛选。