Differentiation of liver progenitor cell line to functional organotypic cultures in 3D nanofibrillar cellulose and hyaluronan-gelatin hydrogels

Abstract

Physiologically relevant hepatic cell culture models must be based on three-dimensional (3D) culture of human cells. However, liver cells are generally cultured in two-dimensional (2D) format that deviates from the normal in vivo morphology. We generated 3D culture environment for HepaRG liver progenitor cells using wood-derived nanofibrillar cellulose (NFC) and hyaluronan-gelatin (HG) hydrogels. Culture of undifferentiated HepaRG cells in NFC and HG hydrogels induced formation of 3D multicellular spheroids with apicobasal polarity and functional bile canaliculi-like structures, structural hallmarks of the liver tissue. Furthermore, hepatobiliary drug transporters, MRP2 and MDR1, were localized on the canalicular membranes of the spheroids and vectorial transport of fluorescent probes towards the biliary compartment was demonstrated. Cell culture in 3D hydrogel supported the mRNA expression of hepatocyte markers (albumin and CYP3A4), and metabolic activity of CYP3A4 in the HepaRG cell cultures. On the contrary, the 3D hydrogel cultures with pre-differentiated HepaRG cells showed decreasing expression of albumin and CYP3A4 transcripts as well as CYP3A4 activity. It is concluded that NFC and HG hydrogels expedite the hepatic differentiation of HepaRG liver progenitor cells better than the standard 2D culture environment. This was shown as improved cell morphology, expression and localization of hepatic markers, metabolic activity and vectorial transport. The NFC and HG hydrogels are promising materials for hepatic cell culture and tissue engineering.

Contextualizing Hepatocyte Functionality of Cryopreserved HepaRG Cell Cultures

“…Recent reports have used HepaRG cells in three-dimensional culture configurations and observed similar increases in metabolic capacity/longevity. However, these models can be complex to generate, highly variable, and difficult to normalize (Malinen et al, 2014;Mueller et al, 2014). Overlaying cryoHepaRG may provide a solution for screening and high-content imaging that obviates the need for confocal microscopy by creating a more coplanar environment.…”

Producing ultrapure wood cellulose nanofibrils and evaluating the cytotoxicity using human skin cells

“…While CNF materials have been compatible with different cell lines (no cytotoxicity) (Alexandrescu, Syverud, Gatti & Chinga-Carrasco, 2013;Bhattacharya et al, 2012;Hua et al, 2014;Lou et al, 2014;Malinen et al, 2014;Tehrani, Nordli, Pukstad, Gethin & Chinga-Carrasco;Vartiainen et al, 2011), a concentration dependent reduction in metabolic activity and/or cell proliferation has also been seen (Čolić, Mihajlović, Mathew, Naseri & Kokol, 2015). TEMPO-mediated oxidation and carboxymethylation are relatively common pre-treatments applied to facilitate the deconstruction of the fibre cell wall and thus the production of CNF (Saito, Nishiyama, Putaux, Vignon & Isogai, 2006;Wågberg, Decher, Norgren, Lindström, Ankerfors & Axnäs, 2008).…”

Highly stable aptamers selected from a 2′-fully modified fGmH RNA library for targeting biomaterials

“…It would not be a large logical leap to assume that such aptamers would also outperform aptamers of lower 2′-modification aptamers and antibody-based affinity molecules that have already been used to functionalize various biosensors, including those on atomic force microscopy, graphene, BLI, and SPR chips [37–40]. fGmH aptamers, specific to a variety of cancer biomarkers, could be used to decorate microfluidic platforms or biomimetic hydrogels for the capture and potential culture of circulating tumor cells (CTCs) derived from a plethora of cancer types, similar to that reported by using EpCAM antibodies [41–43]. fGmH aptamers could also be logically applied to proteome profiling, particularly the highly sensitive single cell detection of cancer cells via a single-cell barcode chip (SCBC) [44].…”