HepaRG Self-Assembled Spheroids in Alginate Beads Meet the Clinical Needs for Bioartificial Liver

Pasqua, Mattia; Pereira, Ulysse; Messina, Antonietta; Lartigue, Claire; Vigneron, Pascale; Dubart‐Kupperschmitt, Anne; Legallais, Cécile

doi:10.1089/ten.tea.2019.0262

Cited by 19 publications

(28 citation statements)

References 36 publications

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“…Compared to primary hepatocytes, HepaRG cell line offers an almost unlimited stock of hepatocytes of constant quality. Because these cells are widely used at two different differentiation stages [18][19][20][21][22][23] , our approach providing a single but versatile microfluidic tool for their culture, differentiation and maturation, is of crucial importance in applications targeting drug screening and toxicity. Rather than adjusting the biological and chemical conditions, additional and innovative physical modifications were made to render the device compatible with the seeding and culture of either proliferative or differentiated HepaRG cells.…”

Section: Discussionmentioning

confidence: 99%

“…This differentiation can be enhanced by adding dimethyl sulfoxide (DMSO) to the culture medium. Both hepatoblast-and hepatocyte-like HepaRG cells are already widely used for liver bioreactor applications 18,19 and to evaluate drug toxicity [20][21][22][23] , respectively. The properties of hepatoblasts and hepatocytes from the HepaRG cell line were evidenced to differ in terms of their migration and adherence: this feature prompted us to adapt the structure and packaging of the chip accordingly, and the culture of these cells under flux for several weeks was then achieved.…”

Section: Introductionmentioning

confidence: 99%

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A Versatile Microfluidic Tool for the 3D Culture of HepaRG Cells Seeded at Various Stages of Differentiation

Boul

Benzoubir

Messina

et al. 2021

Preprint

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The development of livers-on-a-chip aims to provide pharmaceutical companies with reliable systems to perform drug screening and toxicological studies. To that end, microfluidic systems are engineered to mimic the functions and architecture of this organ. In this context we have designed a device that reproduces series of liver microarchitectures, each permitting the 3D culture of hepatocytes by confining them to a chamber that is separated from the medium conveying channel by very thin slits. We modified the structure to ensure its compatibility with the culture of hepatocytes from different sources. Our device was adapted to the migratory and adhesion properties of HepaRG cells at various stages of differentiation. To prevent hepatoblast-like cells from migrating out of the chambers, the slit height was decreased and the medium flow rate increased. Maintaining already differentiated and less adherent cells within the chambers required desensitisation of the system to pressure variations. Using this device, it was possible to keep the cells alive for more than 14 days, during which they achieved a 3D organisation and acquired or maintained their differentiation into hepatocytes. With its multiple micro-chambers for hepatocyte culture, this microfluidic device architecture offers a promising opportunity to provide new tools for drug screening applications.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A Versatile Microfluidic Tool for the 3D Culture of HepaRG Cells Seeded at Various Stages of Differentiation

Boul

Benzoubir

Messina

et al. 2021

Preprint

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“…We recently demonstrated the benefits of using HepaRG cells, capable of differentiating into hepatocyte-like cells (Cerec et al, 2007), encapsulated in alginate beads as the biomass for BAL (Pasqua et al, 2020). The present study, therefore, focused on the transfer of this biomass within a bioreactor compatible with BAL therapy.…”

Section: Discussionmentioning

confidence: 99%

“…The activity of the cytochrome P450-1A1/2 (CYP1A1/2) was studied with the EROD assay as previously described (Diaz, 2000;Pasqua et al, 2020). The microbeads were incubated in ethoxyresorufin (10 μM), prepared in WE, for 1 h at 37°C.…”

Section: Ethoxyresorufin-o-deethylase (Erod) Assaymentioning

confidence: 99%

“…Nevertheless, direct contact and shear stress may be harmful to the cells. Encapsulation of the biomass in semi‐permeable hollow fibres or porous beads is thus often adopted so as to protect it from the hydrodynamic shear stress imposed by the flux (Rebelo et al, 2015) while simultaneously preserving cell activity (Pasqua et al, 2020), and promoting cell‐cell interaction, thus enhancing hepatic‐specific functions (Elkayam et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

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Preclinical characterization of alginate‐poly‐L‐lysine encapsulated HepaRG for extracorporeal liver supply

Pasqua

Pereira

Lartigue

et al. 2020

Biotech & Bioengineering

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We recently demonstrated that HepaRG cells encapsulated into 1.5% alginate beads are capable of self-assembling into spheroids. They adequately differentiate into hepatocyte-like cells, with hepatic features observed at Day 14 post-encapsulation required for external bioartificial liver applications. Preliminary investigations performed within a bioreactor under shear stress conditions and using a culture medium mimicking acute liver failure (ALF) highlighted the need to reinforce beads with a polymer coating. We demonstrated in a first step that a poly-L-lysine coating improved the mechanical stability, without altering the metabolic activities necessary for bioartificial liver applications (such as ammonia and lactate elimination). In a second step, we tested the optimized biomass in a newly designed perfused dynamic bioreactor, in the presence of the medium model for pathological plasma for 6 h. Performances of the biomass were enhanced as compared to the steady configuration, demonstrating its efficacy in decreasing the typical toxins of ALF. This type of bioreactor is easy to scale up as it relies on the number of micro-encapsulated cells, and could provide an adequate hepatic biomass for liver supply. Its design allows it to be integrated into a hybrid artificial/bioartificial liver setup for further clinical studies regarding its impact on ALF animal models.

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Advanced Techniques and Awaited Clinical Applications for Human Pluripotent Stem Cell Differentiation into Hepatocytes

et al. 2021

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Liver transplantation is currently the only curative treatment for several liver diseases such as acute liver failure, end-stage liver disorders, primary liver cancers and certain genetic conditions.Unfortunately, despite improvements to transplantation techniques, including live donor transplantation, the number of organs available remains insufficient to meet patient needs.Hepatocyte transplantation has enabled some encouraging results as an alternative to organ transplantation, but primary hepatocytes are little available and cannot be amplified using traditional 2D culture systems. Indeed, although recent studies have tended to show that 3D culture enables long term hepatocyte culture, it is still agreed that like most adult primary cell types, they remain refractory to in vitro expansion. Because of their exceptional properties, human pluripotent stem cells (hPSCs) can be amplified indefinitely and differentiated into any cell type, including liver cells. While many teams have worked on hepatocyte differentiation, there has been a consensus that cells obtained after hPSC differentiation have more foetal than adult hepatocyte characteristics. New technologies have been used to improve the differentiation process in recent years. This review discusses the technical improvements made to hepatocyte differentiation protocols and the clinical approaches developed to date and anticipated in the near future.

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HepaRG Self-Assembled Spheroids in Alginate Beads Meet the Clinical Needs for Bioartificial Liver

Cited by 19 publications

References 36 publications

A Versatile Microfluidic Tool for the 3D Culture of HepaRG Cells Seeded at Various Stages of Differentiation

A Versatile Microfluidic Tool for the 3D Culture of HepaRG Cells Seeded at Various Stages of Differentiation

Preclinical characterization of alginate‐poly‐L‐lysine encapsulated HepaRG for extracorporeal liver supply

Advanced Techniques and Awaited Clinical Applications for Human Pluripotent Stem Cell Differentiation into Hepatocytes

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