3D culture of HepaRG cells in GelMa and its application to bioprinting of a multicellular hepatic model. (February 2021)

• Record Type: Journal Article
• Authors: Cuvellier, Marie; Ezan, Frédéric; Oliveira, Hugo; Rose, Sophie; Fricain, Jean-Christophe; Langouët, Sophie; Legagneux, Vincent; Baffet, Georges
• Is Part Of: Biomaterials. Volume 269(2021)
• Publication Date: 2021-02
• Subjects: Bioprinting -- Methacrylated gelatin -- Hepatocyte -- HepaRG -- 3D liver models; 2D two-dimensions -- 3D three-dimensions -- 3 MC 3-methylcholanthrene -- CYP Cytochromes P450 -- CDDP cisplatine -- DMSO dimethyl sulfoxide -- ECM extracellular matrix -- EdU 5-ethynyl-2′-deoxyuridine -- EROD Ethoxyresorufin-O-deethylation GelMa -- methacrylated gelatin HSC -- hepatic stellate cells LAP -- lithium phenyl-2, 4, 6 trimethylbenzoylphosphinate MROD -- Methyresorufin-O-deethylation TGFβ-1 -- Transforming growth factor beta 1 TPEF -- two-photon excitation microscopy SHG second harmonic generation; Biomedical materials -- Periodicals; Biocompatible Materials -- Periodicals; Biomatériaux -- Périodiques; 610.28
• DOI: 10.1016/j.biomaterials.2020.120611
• Languages: English
• ISSNs: 0142-9612

Abstract

Abstract: Bioprinting is an emergent technology that has already demonstrated the capacity to create complex and/or vascularized multicellular structures with defined and organized architectures, in a reproducible and high throughput way. Here, we present the implementation of a complex liver model by the development of a three-dimensional extrusion bioprinting process, including parameters for matrix polymerization of methacrylated gelatin, using two hepatic cell lines, Huh7 and HepaRG. The printed structures exhibited long-term viability (28 days), proliferative ability, a relevant hepatocyte phenotype and functions equivalent to or better than those of their 2D counterparts using standard DMSO treatment. This work served as a basis for the bioprinting of complex multicellular models associating the hepatic parenchymal cells, HepaRG, with stellate cells (LX-2) and endothelial cells (HUVECs), able of colonizing the surface of the structure and thus recreating a pseudo endothelial barrier. When bioprinted in 3D monocultures, LX-2 expression was modulated by TGFβ-1 toward the induction of myofibroblastic genes such as ACTA2 and COL1A1. In 3D multicellular bioprinted structures comprising HepaRG, LX-2 and endothelial cells, we evidenced fibrillar collagen deposition, which is never observed in monocultures of either HepaRG or LX-2 alone. These observations indicate that a precise control of cellular communication is required to recapitulate key steps of fibrogenesis. Bioprinted 3D co-cultures therefore open up new perspectives in studying the molecular and cellular basis of fibrosis development and provide better access to potential inducers and inhibitors of collagen expression and deposition. Graphical abstract: Image 1