Alteration of pancreatic carcinoma and promyeloblastic cell adhesion in liver microvasculature by co-culture of hepatocytes, hepatic stellate cells and endothelial cells in a physiologically-relevant model

Abstract

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.

Hydrogel microfluidic‐based liver‐on‐a‐chip: Mimicking the mass transfer and structural features of liver

“…Although the distance between inner and outer channels of 1 mm was much larger than the space of Disse in the liver, which has an extremely small thickness of 0.2–1 µm (Kang et al, 2015), the permeable F127‐DA hydrogel still facilitated the interaction between endothelial and hepatic cells via diffusion of soluble factors (e.g., VEGF and hepatocyte growth factor). Such factors generated by endothelial cells could benefit the expression of liver‐specific functions by hepatocytes (Danoy et al, 2017; Ding et al, 2010).…”

Protective effects of specific cannabinoid receptor 2 agonist GW405833 on concanavalin A-induced acute liver injury in mice

“…To explore why activation of CB2R in T cells could protect hepatocytes against acute injury to the liver, a coculture cell model was employed as previously described [29,30]. Addition of GW inhibited the Con A-induced apoptosis of L02, and AM630 reversed this effect.…”

Engineered basement membranes: fromin vivoconsiderations to cell-based assays

“…To study cancer progression from endothelial-mesenchymal transition (EMT) to the cancer cell adhesion, invasion and vascularisation, various in vitro cancer models have been developed. 77,78,140,153,154 Nevertheless, the generation of a good advanced cell-based assay to model cancer still remains challenging as it depends on both biophysical and biochemical properties of BMs. Kamm's group at the Massachusetts Institute of Technology, has been developing advanced cell-based assays to study cancer progression.…”