Tissue-Engineered Bile Ducts for Disease Modeling and Therapy

Wang, Zhenguo; Faria, João; Penning, Louis C.; Masereeuw, Rosalinde; Spee, Bart

doi:10.1089/ten.tec.2020.0283

Cited by 11 publications

(8 citation statements)

References 167 publications

(190 reference statements)

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“…Recently, synthetic and natural materials have been used to construct bioengineered bile ducts for transplantation purposes (Wang et al, 2021). Sampaziotis et al (2017b) described collagen scaffolds populated by human extrahepatic organoids (ECOs) that formed a bioengineered bile duct, which were able to replace the native mouse extrahepatic bile duct.…”

Section: Discussionmentioning

confidence: 99%

“…Epithelial markers, tight junction protein 1 (TJP1, also known as ZO1), and E-cadherin 1 (CDH1), as well as cholangiocyte markers K7 and K19, were present in CLCs. More importantly, the CLCs expressed cholangiocyte-specific transporters, secretin receptor (SCTR), and TGR5 (also known as GPBAR1, encoded by GPBAR1) that play an important role in sensing and signaling and appear in mature cholangiocytes (Guo et al, 2016;Sato et al, 2018;Wang et al, 2021). In addition, the CLCs showed F-actin indicated by the staining for phalloidin, as a marker for the apical domains which demonstrates the CLCs developed apicobasal epithelial polarity, with the apical surface facing the lumen.…”

Section: In Vitro Generation Of Clcs From Icosmentioning

confidence: 96%

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Human Cholangiocytes Form a Polarized and Functional Bile Duct on Hollow Fiber Membranes

Wang

Faria

Laan

et al. 2022

Front. Bioeng. Biotechnol.

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Liver diseases affect hundreds of millions of people worldwide; most often the hepatocytes or cholangiocytes are damaged. Diseases of the biliary tract cause severe patient burden, and cholangiocytes, the cells lining the biliary tract, are sensitive to numerous drugs. Therefore, investigations into proper cholangiocyte functions are of utmost importance, which is restricted, in vitro, by the lack of primary human cholangiocytes allowing such screening. To investigate biliary function, including transepithelial transport, cholangiocytes must be cultured as three-dimensional (3D) ductular structures. We previously established murine intrahepatic cholangiocyte organoid-derived cholangiocyte-like cells (CLCs) and cultured them onto polyethersulfone hollow fiber membranes (HFMs) to generate 3D duct structures that resemble native bile ducts at the structural and functional level. Here, we established an efficient, stepwise method for directed differentiation of human intrahepatic cholangiocyte organoids (ICOs) into CLCs. Human ICO-derived CLCs showed key characteristics of cholangiocytes, such as the expression of structural and functional markers, formation of primary cilia, and P-glycoprotein-mediated transport in a polarized fashion. The organoid cultures exhibit farnesoid X receptor (FXR)-dependent functions that are vital to liver bile acid homeostasis in vivo. Furthermore, human ICO-derived CLCs cultured on HFMs in a differentiation medium form tubular architecture with some tight, confluent, and polarized monolayers that better mimic native bile duct characteristics than differentiated cultures in standard 2D or Matrigel-based 3D culture plates. Together, our optimized differentiation protocol to obtain CLC organoids, when applied on HFMs to form bioengineered bile ducts, will facilitate studying cholangiopathies and allow developing therapeutic strategies.

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Section: Discussionmentioning

confidence: 99%

Section: In Vitro Generation Of Clcs From Icosmentioning

confidence: 96%

Human Cholangiocytes Form a Polarized and Functional Bile Duct on Hollow Fiber Membranes

Wang

Faria

Laan

et al. 2022

Front. Bioeng. Biotechnol.

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“…BA secretion) and toxicological studies. For this, the biliary epithelial cell lines, H69, MMNK-1 and HepaRG presented key biliary markers and have been used to mimic the cholangiocyte phenotype [31]. Furthermore, primary cholangiocytes possess the morphological and functional characteristics of the original tissue, but the primary sources are scarce and rapidly dedifferentiate in (two-dimensional) culture conditions, resulting in the shortage of FXR expression [32] that indicate the loss of speci c functions including BA homeostasis [33].…”

Section: Discussionmentioning

confidence: 99%

Cholangiocyte Organoids to Model Chlorpromazine-Induced Bile Duct Injury

Wang,

Xing,

Laan

et al. 2023

Preprint

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Background Drug induced bile duct injury is a frequently observed clinical problem leading to a wide range of pathological features. During the past decades, several agents have been identified with various postulated mechanisms of bile duct damage, however, mostly still poorly understood. Methods Here, we investigated the mechanisms of chlorpromazine(CPZ) induced bile duct injury using advanced in vitro cholangiocyte cultures. Intrahepatic cholangiocyte organoids (ICOs) were driven into mature cholangiocyte like cells (CLCs), which were exposed to CPZ under cholestatic or non-cholestatic conditions through the addition of a bile acid cocktail. Results CPZ caused loss of monolayer integrity by reducing expression levels of tight junction protein 1 (TJP1), E-cadherin 1 (CDH1) and lysyl oxidase homolog 2 (LOXL2). Loss of zonula occuludens-1 (ZO-1) and E-cadherin was confirmed by immunostaining after exposure to CPZ and rhodamine-123 leakage further confirmed disruption of the cholangiocyte barrier function. Furthermore, oxidative stress seemed to play a major role in the early damage response by CPZ. The drug also decreased expression of three main basolateral bile acid transporters, ABCC3 (ATP binding cassette subfamily C member 3), SLC51A/B (solute carrier family 51 subunit alpha/beta) and multidrug resistance transporter ABCB1 (ATP binding cassette subfamily B member 1), thereby contributing to bile acid accumulation. CPZ did not induce an inflammatory response by itself, but addition of TNFα revealed a synergistic effect. Conclusion These results show that ICOs present a model to identify toxic drugs affecting the bile ducts while providing mechanistic insights into hepatotoxicity.

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“…On the contrary, few protocols have been reported to induce cholangiocyte‐like cells. [ 6 , 9 , 43 ] Cholangiocytes share the same germ layer as hepatocytes; therefore, the definitive endoderm was induced using the same initial steps as the hepatocyte protocol. [ 2 , 8 ] Cholangiocyte specification and maturation was done by treating the cells with several growth factors and molecules, TGFβ1 and sDLL‐1.…”

Section: Discussionmentioning

confidence: 99%

Transmembrane channel activity in human hepatocytes and cholangiocytes derived from induced pluripotent stem cells

Florentino

Coard

et al. 2022

Hepatol Commun

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The initial creation of human-induced pluripotent stem cells (iPSCs) set the foundation for the future of regenerative medicine. Human iPSCs can be differentiated into a variety of cell types in order to study normal and pathological molecular mechanisms. Currently, there are well-defined protocols for the differentiation, characterization, and establishment of functionality in human iPSC-derived hepatocytes (iHep) and iPSC-derived cholangiocytes (iCho). Electrophysiological study on chloride ion efflux channel activity in iHep and iCho cells has not been previously reported. We generated iHep and iCho cells and characterized them based on hepatocyte-specific and cholangiocyte-specific markers. The relevant transmembrane channels were selected: cystic fibrosis transmembrane conductance regulator, leucine rich repeat-containing 8 subunit A, and transmembrane member 16 subunit A. To measure the activity in these channels, we used whole-cell patch-clamp techniques with a standard intracellular and extracellular solution. Our iHep and iCho cells demonstrated definitive activity in the selected transmembrane channels, and this approach may become an important tool for investigating human liver biology of cholestatic diseases.

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Tissue-Engineered Bile Ducts for Disease Modeling and Therapy

Cited by 11 publications

References 167 publications

Human Cholangiocytes Form a Polarized and Functional Bile Duct on Hollow Fiber Membranes

Human Cholangiocytes Form a Polarized and Functional Bile Duct on Hollow Fiber Membranes

Cholangiocyte Organoids to Model Chlorpromazine-Induced Bile Duct Injury

Transmembrane channel activity in human hepatocytes and cholangiocytes derived from induced pluripotent stem cells

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