Self‐assembled liver organoids recapitulate hepatobiliary organogenesis in vitro

Vyas, Dipen; Baptista, Pedro M.; Brovold, Matthew; Moran, Emma; Gaston, Brandon; Booth, Chris; Samuel, Michael S.; Atala, Anthony; Söker, Shay

doi:10.1002/hep.29483

Cited by 100 publications

(100 citation statements)

References 40 publications

Supporting

Mentioning

100

Contrasting

Order By: Relevance

“…Isolated perfused intrahepatic bile duct units maintain their in vivo organization and have been used effectively to characterize cholangiocyte cilia as mechanical and osmotic sensors, but they do not permit measurement of monolayer permeability, manipulation of the underlying matrix, or incorporation of genetically modified or other specific populations of cholangiocytes . Cholangiocytes in 3D spheroid culture differentiate well, can be cultured in physiological matrices, and demonstrate barrier functions and transport activities . However, spheroids are highly variable in shape and size, and it is difficult to control their positions in a gel (for microscopy), sample lumen content, or treat the cholangiocyte apical surface.…”

Section: Discussionmentioning

confidence: 99%

A Bile Duct‐on‐a‐Chip With Organ‐Level Functions

Khandekar

Llewellyn

et al. 2019

Hepatology

View full text Add to dashboard Cite

Background and Aims Chronic cholestatic liver diseases, such as primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC), are frequently associated with damage to the barrier function of the biliary epithelium. Here, we report on a bile duct‐on‐a‐chip that phenocopies not only the tubular architecture of the bile duct in three dimensions, but also its barrier functions. Approach and Results We showed that mouse cholangiocytes in the channel of the device became polarized and formed mature tight junctions, that the permeability of the cholangiocyte monolayer was comparable to ex vivo measurements, and that cholangiocytes in the device were mechanosensitive (as demonstrated by changes in calcium flux under applied luminal flow). Permeability decreased significantly when cells formed a compact monolayer with cell densities comparable to those observed in vivo. This device enabled independent access to the apical and basolateral surfaces of the cholangiocyte channel, allowing proof‐of‐concept toxicity studies with the biliary toxin, biliatresone, and the bile acid, glycochenodeoxycholic acid. The cholangiocyte basolateral side was more vulnerable than the apical side to treatment with either agent, suggesting a protective adaptation of the apical surface that is normally exposed to bile. Further studies revealed a protective role of the cholangiocyte apical glycocalyx, wherein disruption of the glycocalyx with neuraminidase increased the permeability of the cholangiocyte monolayer after treatment with glycochenodeoxycholic acid. Conclusions This bile duct‐on‐a‐chip captured essential features of a simplified bile duct in structure and organ‐level functions and represents an in vitro platform to study the pathophysiology of the bile duct using cholangiocytes from a variety of sources.

show abstract

Section: Discussionmentioning

confidence: 99%

A Bile Duct‐on‐a‐Chip With Organ‐Level Functions

Khandekar

Llewellyn

et al. 2019

Hepatology

View full text Add to dashboard Cite

show abstract

“…The seeding procedure was performed as previously described (Vyas et al, ). Briefly, 2.5 × 10 5 iHeps were suspended in 10 μl iHep medium for each disk.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, the decellularized liver tissue may be a better alternative. Previously, we and others have reported that decellularized liver tissue could provide an excellent environment for the in vitro differentiation of hepatic stem cells (Vyas et al, ; Wang et al, ) as well as maintenance of primary hepatocytes (Soto‐Gutierrez et al, ). However, it is still unknown whether iHeps can repopulate decellularized liver tissue and whether the maturation of iHeps can be enhanced by this more natural environment.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Hepatocyte‐like cells generated by direct reprogramming from murine somatic cells can repopulate decellularized livers

Chen

Plá-Palacín

Baptista

et al. 2018

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

Direct reprogramming represents an easy technique to generate induced hepatocytes (iHeps) from somatic cells. However, current protocols are accompanied by several drawbacks as iHeps are heterogenous and lack fully mature phenotypes of primary hepatocytes. Here, we established a polycistronic expression system to induce the direct reprogramming of mouse embryonic fibroblasts towards hepatocytes. The resulting iHeps are homogenous and display key properties of primary hepatocytes, such as expression of hepatocyte markers, albumin secretion, and presence of liver transaminases. iHeps also possess the capacity to repopulate decellularized liver tissue and exhibit enhanced hepatic maturation. As such, we present a novel strategy to generate homogenous and functional iHeps for applications in tissue engineering and cell therapy.

show abstract

“…These 3D liver organoids recapitulated several aspects of hepatobiliary organogenesis and resulted in concomitant formation of progressively more differentiated hepatocytes and bile duct structures. 103 Applications of the ECM-based 3D culture are limited by the mass transfer barriers formed by the top layer, batch-to-batch variation of ECM, and uncontrollable coating.…”

Section: Strategies To Promote Hepatocyte Differentiation and Maturationmentioning

confidence: 99%

Biotechnology Challenges to In Vitro Maturation of Hepatic Stem Cells

et al. 2018

Self Cite

View full text Add to dashboard Cite

The incidence of liver disease is increasing globally. The only curative therapy for severe end-stage liver disease, liver transplantation, is limited by the shortage of organ donors. In vitro models of liver physiology have been developed and new technologies and approaches are progressing rapidly. Stem cells might be used as a source of liver tissue for development of models, therapies, and tissue-engineering applications. However, we have been unable to generate and maintain stable and mature adult liver cells ex vivo. We review factors that promote hepatocyte differentiation and maturation, including growth factors, transcription factors, microRNAs, small molecules, and the microenvironment. We discuss how the hepatic circulation, microbiome, and nutrition affect liver function, and the criteria for considering cells derived from stem cells to be fully mature hepatocytes. We explain the challenges to cell transplantation and consider future technologies for use in hepatic stem cell maturation, including 3-dimensional biofabrication and genome modification.

show abstract

Self‐assembled liver organoids recapitulate hepatobiliary organogenesis in vitro

Cited by 100 publications

References 40 publications

A Bile Duct‐on‐a‐Chip With Organ‐Level Functions

A Bile Duct‐on‐a‐Chip With Organ‐Level Functions

Hepatocyte‐like cells generated by direct reprogramming from murine somatic cells can repopulate decellularized livers

Biotechnology Challenges to In Vitro Maturation of Hepatic Stem Cells

Contact Info

Product

Resources

About