In vitro models of the liver microvasculature, especially with respect to cancer cell extravasation, should include not only endothelial and cancer cells but also surrounding cells to mimic the physiological situation. To this end, in the present study, we established a physiologically-relevant hierarchical co-culture model by stacking layers of primary rat hepatocytes (Hep), hepatic stellate cells embedded in collagen gel (LX-2) and endothelial cells (HUVECs) on a specially designed oxygen-permeable polydimethylsiloxane PDMS bottom plate. The model was used to investigate the role and contribution of each of the three cell types in pancreatic cancer and promyeloblast cell adhesion. In particular, we showed an increase in albumin production by the primary hepatocytes and in the consumption of the produced vascular endothelial growth factors (VEGFs). Furthermore, in co-culture, the HUVECs exhibited a mature vascular endothelial and non-inflamed phenotype, as evidenced by Stabilin-1, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), intercellular adhesion molecule (ICAM-1), and vascular adhesion protein-1 (VAP-1) expression. The HUVECs were also successfully activated with an inflammatory cytokine and their ICAM-1 response was found to be higher in monoculture compared to co-culture. Additionally, the adhesion of MiaPaCa-2 pancreatic cancer cells and HL60 promyeloblasts was tested in both cases (i.e.: activation or not by an inflammatory cytokine). It has been found that their adhesion was always reduced in the co-culture model. These results highlight the importance of integrating hepatic stellate cells in the design of biomimetic models of the hepatic endothelial barrier.

中文翻译：

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在生理相关模型中，通过肝细胞，肝星状细胞和内皮细胞的共培养，肝脏微脉管系统中胰腺癌和早幼粒细胞粘附的改变。

肝微脉管系统的体外模型，尤其是关于癌细胞外渗的模型，不仅应包括内皮细胞和癌细胞，而且还应包括周围细胞以模拟生理状况。为此，在本研究中，我们通过堆叠原代大鼠肝细胞（Hep），包埋在胶原蛋白凝胶（LX-2）中的肝星状细胞和内皮细胞（HUVEC）的层，建立了生理相关的分层共培养模型。专门设计的透氧聚二甲基硅氧烷PDMS底板。该模型用于研究三种细胞类型在胰腺癌和早幼粒细胞粘附中的作用和贡献。特别是，我们显示出原代肝细胞的白蛋白产生增加以及所产生的血管内皮生长因子（VEGF）的消耗增加。此外，在共培养中，HUVEC表现出成熟的血管内皮和非炎症表型，如Stabilin-1，淋巴管内皮透明质酸受体1（LYVE-1），细胞间粘附分子（ICAM-1）和血管粘附蛋白1（VAP-1）的表达。HUVEC也已被炎性细胞因子成功激活，并且与共培养相比，在单培养中它们的ICAM-1反应更高。另外，在两种情况下都测试了MiaPaCa-2胰腺癌细胞和HL60早幼粒细胞的粘附性（即：是否通过炎性细胞因子激活）。已经发现在共培养模型中它们的粘附力总是降低的。