scholarly journals Cholangiocyte organoids from human bile retain a local phenotype and can repopulate bile ducts in vitro

2021 ◽  
Vol 11 (12) ◽  
Author(s):  
Floris J. M. Roos ◽  
Haoyu Wu ◽  
Jorke Willemse ◽  
Ruby Lieshout ◽  
Laura A. Muñoz Albarinos ◽  
...  
Keyword(s):  
Bile Ducts ◽  
Human Bile

Adenosine nucleotides in bile

1996 ◽  
Vol 270 (2) ◽  
pp. G246-G252 ◽  
Author(s):  
R. S. Chari ◽  
S. M. Schutz ◽  
J. E. Haebig ◽  
G. H. Shimokura ◽  
P. B. Cotton ◽  
...  
Keyword(s):  
Purinergic Receptors ◽  
Purinergic Receptor ◽  
Receptor Activation ◽  
Hepatoma Cells ◽  
Human Bile ◽  
Nucleotide Release ◽  
Cholangiocarcinoma Cell ◽  
Adenosine Nucleotides ◽  
Vitro Model

Activation of purinergic receptors by ATP stimulates Cl- efflux in biliary epithelial cells. To determine whether purinergic agonists are present under physiological conditions, we have assayed mammalian bile for nucleotides and assessed whether hepatoma and cholangiocarcinoma cell lines are capable of nucleotide release. Bile samples were collected from human, rat, and pig donors and assayed for nucleotide concentrations by luminometry. ATP, ADP, and AMP were present in bile from each species, and the average total nucleotide concentration in human bile was 5.21 +/- 0.91 microM (n = 16). In an in vitro model of HTC rat hepatoma cells or Mz-ChA-1 cholangiocarcinoma cells on a superfused column, nucleotides were present in the effluent from each cell type. Addition of alpha, beta-methyleneadenosine 5'-diphosphate (50 microM) to inhibit 5'-nucleotidase activity increased AMP concentrations two- to threefold. Exposure to forskolin (100 microM) or ionomycin (2 microM) stimulated nucleotide release from cholangiocarcinoma but not hepatoma cells. These studies indicate that adenosine nucleotides are present in bile in concentrations sufficient to activate purinergic receptors. Purinergic receptor activation by local nucleotide release might constitute an autocrine and/or paracrine mechanism for modulation of biliary secretion.

scholarly journals Salmonella L-forms: formation in human bile in vitro and isolation culture from patients' gallbladder samples by a non-high osmotic isolation technique

2015 ◽  
Vol 21 (5) ◽  
pp. 470.e9-470.e16
Author(s):  
D.N. Wang ◽  
W.J. Wu ◽  
T. Wang ◽  
Y.Z. Pan ◽  
K.L. Tang ◽  
...  
Keyword(s):  
Isolation Technique ◽  
Human Bile

scholarly journals Cell isolation and Organoid Culture from Mouse Liver and Small Intestine

2021 ◽  
Author(s):  
Wenyi Chen ◽  
Qigu Yao ◽  
Ruo Wang ◽  
Yanping Xu ◽  
Jiong Yu ◽  
...  
Keyword(s):  
Small Intestine ◽  
Bile Ducts ◽  
Disease Modeling ◽  
Organoid Culture ◽  
Intrahepatic Bile Ducts ◽  
Tissue Organization ◽  
Intestinal Organoids ◽  
Gravity Settling ◽  
Small Intestinal

Abstract Background: Organoid culture enables disease modeling and drug screening in vitro. Organoids are from organs (e.g., brain, small intestine, kidney, lung, and liver). To facilitate the establishment of liver and small-intestinal organoids, we developed a protocol for collecting cholangiocytes and crypts and culturing organoids.Methods: Cholangiocytes were collected from intrahepatic bile ducts, gallbladder, and crypts from the small intestine using gravity settling and multi-step centrifugation methods, and embedded in Matrigel to grow into three-dimensional spheroids in suitable culture medium. Passaging, cryopreservation, and thawing were performed to assess organoid cell stability. RNA and DNA extraction, as well as immunostaining procedure were optimized. For preclinical modeling, the growth rate of cholangiocyte organoids (cho-orgs) was harmonized.Results: Large amount of Cholangiocytes and small intestine crypts were collected. Cholangiocytes developed into cyst-like structures after 3–4 days in Matrigel. After culture for 1–2 weeks, small-intestinal organoids developed buds and formed a mature structure. Cho-orgs from intrahepatic bile ducts grew more slowly but were longer lasting, expressed the cholangiocyte markers Krt19 and Krt7, and recapitulated the in vivo tissue organization.Conclusions: The protocol takes 2–4 weeks to establish a stable organoid growth system. Organoids could be stably passaged, cryopreserved, and recovered. The organoids retained tissue characteristics, including marker expression.

scholarly journals Tenascin-W Is a Novel Stromal Marker in Biliary Tract Cancers

2021 ◽  
Vol 11 ◽  
Author(s):  
Ismaïl Hendaoui ◽  
Ahlem Lahmar ◽  
Luca Campo ◽  
Sihem Mebarki ◽  
Sandrine Bichet ◽  
...  
Keyword(s):  
Biliary Tract ◽  
Stromal Cells ◽  
Poor Prognosis ◽  
Bile Ducts ◽  
Tumor Stroma ◽  
Biliary Tract Cancers ◽  
Tenascin C ◽  
Normal Tissues ◽  
Biliary Tract Tumors

Extrahepatic cancers of the biliary system are typically asymptomatic until after metastasis, which contributes to their poor prognosis. Here we examined intrahepatic cholangiocarcinomas (n = 8), carcinomas of perihilar bile ducts (n = 7), carcinomas of the gallbladder (n = 11) and hepatic metastasis from carcinomas of the gallbladder (n = 4) for the expression of the extracellular matrix glycoproteins tenascin-C and tenascin-W. Anti-tenascin-C and anti-tenascin-W immunoreactivity was found in all biliary tract tumors examined. Unlike tenascin-C, tenascin-W was not detected in normal hepatobiliary tissue. Tenascin-W was also expressed by the cholangiocarcinoma-derived cell line Huh-28. However, co-culture of Huh-28 cells with immortalized bone marrow-derived stromal cells was necessary for the formation and organization of tenascin-W fibrils in vitro. Our results indicate that tenascin-W may be a novel marker of hepatobiliary tumor stroma, and its absence from many normal tissues suggests that it may be a potential target for biotherapies.

scholarly journals Cultivation and Genomic Characterization of the Bile Bacterial Species From Cholecystitis Patients

2021 ◽  
Vol 12 ◽  
Author(s):  
Qiulong Yan ◽  
Siyi Zhang ◽  
Shenghui Li ◽  
Guangyang Wang ◽  
Aiqin Zhang ◽  
...  
Keyword(s):  
Bile Acids ◽  
Bacterial Species ◽  
Fluoroquinolone Resistance ◽  
Metagenomic Sequencing ◽  
Multidrug Resistance Proteins ◽  
Aminoglycoside Resistance ◽  
Human Bile ◽  
Resistance Proteins ◽  
Bile Bacteria

The microbes in human bile are closely related to gallbladder health and other potential disorders. Although the bile microbial community has been investigated by recent studies using amplicon or metagenomic sequencing technologies, the genomic information of the microbial species resident in bile is rarely reported. Herein, we isolated 138 bacterial colonies from the fresh bile specimens of four cholecystitis patients using a culturome approach and genomically characterized 35 non-redundant strains using whole-genome shotgun sequencing. The bile bacterial isolates spanned 3 classes, 6 orders, 10 families, and 14 genera, of which the members of Enterococcus, Escherichia–Shigella, Lysinibacillus, and Enterobacter frequently appeared. Genomic analysis identified three species, including Providencia sp. D135, Psychrobacter sp. D093, and Vibrio sp. D074, which are not represented in existing reference genome databases. Based on the genome data, the functional capacity between bile and gut isolates was compared. The bile strains encoded 5,488 KEGG orthologs, of which 4.9% were specific to the gut strains, including the enzymes involved in biofilm formation, two-component systems, and quorum-sensing pathways. A total of 472 antibiotic resistance genes (ARGs) were identified from the bile genomes including multidrug resistance proteins (42.6%), fluoroquinolone resistance proteins (12.3%), aminoglycoside resistance proteins (9.1%), and β-lactamase (7.2%). Moreover, in vitro experiments showed that some bile bacteria have the capabilities for bile salt deconjugation or biotransformation (of primary bile acids into secondary bile acids). Although the physiological or pathological significance of these bacteria needs further exploration, our works expanded knowledge about the genome, diversity, and function of human bile bacteria.

Organoids regenerate human bile ducts

Science ◽  
2021 ◽  
Vol 371 (6531) ◽  
pp. 793.19-795
Author(s):  
Priscilla N. Kelly
Keyword(s):  
Bile Ducts ◽  
Human Bile
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