scholarly journals Generation of mouse hepatobiliary organoids from hepatocyte progenitors and cholangiocytes isolated from healthy adult mouse liver

2021 ◽  
Author(s):  
Naoki Tanimizu ◽  
Norihisa Ichinohe ◽  
Yasushi Sasaki ◽  
Tohru Itoh ◽  
Ryo Sudo ◽  
...  
Keyword(s):  
Adult Mouse ◽  
Functional Connection ◽  
Report Generation ◽  
Mouse Hepatocyte ◽  
Functional Connections ◽  
Metabolic Functions ◽  
Small Hepatocytes ◽  
Liver Organoids ◽  
Adult Mouse Liver

Abstract Number of liver organoids have been reported, though it is not clearly shown whether the functional connection between hepatocytes and cholangiocytes is recapitulated in those organoids. Here, we report generation of a hepatobiliary tubular organoid (HBTO) using mouse hepatocyte progenitors called small hepatocytes (SHs) and cholangiocytes. SHs differentiate and form the bile canalicular network in HBTOs and secret metabolites into the canaliculi, which are then transported into the biliary structure. Hepatocytes in the organoid acquire and maintain metabolic functions including albumin secretion and cytochrome P450 activities, over the long term. We provide the step-by-step protocol for induction of HBTO including isolation of cholangiocytes and SHs and co-culture of these two types of cell to generate functional connections between hepatocytes and cholangiocytes.

scholarly journals Generation of functional liver organoids on combining hepatocytes and cholangiocytes with hepatobiliary connections ex vivo

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Naoki Tanimizu ◽  
Norihisa Ichinohe ◽  
Yasushi Sasaki ◽  
Tohru Itoh ◽  
Ryo Sudo ◽  
...  
Keyword(s):  
Liver Tissue ◽  
Ex Vivo ◽  
Drainage System ◽  
Functional Connection ◽  
Functional Connections ◽  
Intrahepatic Bile Ducts ◽  
Metabolic Functions ◽  
Bile Drainage ◽  
Liver Organoids

AbstractIn the liver, the bile canaliculi of hepatocytes are connected to intrahepatic bile ducts lined with cholangiocytes, which remove cytotoxic bile from the liver tissue. Although liver organoids have been reported, it is not clear whether the functional connection between hepatocytes and cholangiocytes is recapitulated in those organoids. Here, we report the generation of a hepatobiliary tubular organoid (HBTO) using mouse hepatocyte progenitors and cholangiocytes. Hepatocytes form the bile canalicular network and secrete metabolites into the canaliculi, which are then transported into the biliary tubular structure. Hepatocytes in HBTO acquire and maintain metabolic functions including albumin secretion and cytochrome P450 activities, over the long term. In this study, we establish functional liver tissue incorporating a bile drainage system ex vivo. HBTO enable us to reproduce the transport of hepatocyte metabolites in liver tissue, and to investigate the way in which the two types of epithelial cells establish functional connections.

scholarly journals Generation of functional liver tissue by establishing hepatobiliary connections ex vivo

2020 ◽  
Author(s):  
Naoki Tanimizu ◽  
Norihisa Ichinohe ◽  
Yasushi Sasaki ◽  
Tohru Itoh ◽  
Ryo Sudo ◽  
...  
Keyword(s):  
Liver Tissue ◽  
Ex Vivo ◽  
Drainage System ◽  
Bile Canaliculi ◽  
Mouse Hepatocyte ◽  
Functional Connections ◽  
Intrahepatic Bile Ducts ◽  
Metabolic Functions ◽  
Bile Drainage

Abstract In the liver, the bile canaliculi of hepatocytes are connected to intrahepatic bile ducts lined with cholangiocytes, which remove cytotoxic bile from the liver tissue. We have developed a hepatobiliary organoid using mouse hepatocyte progenitors and cholangiocytes. Hepatocyte metabolites were secreted into the bile canaliculi, and then transported into the biliary structure. Hepatocytes in the organoid acquired and maintained metabolic functions including albumin secretion and cytochrome P450 activities, over the long term. In this study, we established functional liver tissue incorporating a bile drainage system ex vivo. This hepatobiliary organoid enabled us to reproduce the transport of hepatocyte metabolites in liver tissue, and to investigate the way in which the two types of epithelial cells establish functional connections.

scholarly journals Hepatocyte-specific deletion of XBP1 sensitizes mice to liver injury through hyperactivation of IRE1α

2020 ◽  
Author(s):  
Caroline C. Duwaerts ◽  
Kevin Siao ◽  
Russell K. Soon ◽  
Chris Her ◽  
Takao Iwawaki ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Liver Injury ◽  
Er Stress ◽  
Adult Mouse ◽  
Active Form ◽  
Cellular Responses ◽  
Genetic Deletion ◽  
Specific Deletion ◽  
Adult Mouse Liver

AbstractX-box binding protein-1 (XBP1) is a transcription factor that plays a central role in controlling cellular responses to endoplasmic reticulum (ER) stress. Under stress conditions, the transcriptionally active form of XBP1 is generated by unique splicing of Xbp1 mRNA by the ER-resident protein inositol-requiring enzyme-1 (IRE1α). Genetic deletion of XBP1 has multiple consequences: some resulting from the loss of the transcription factor per se, and others related to compensatory upstream activation of IRE1α. The objective of the current study was to investigate the effects of XBP1 deletion in adult mouse liver and determine to what extent they are direct or indirect. XBP1 was deleted specifically from hepatocytes in adult Xbp1fl/fl mice using AAV8-Transthyretin-Cre (Xbp1Δhep). Xbp1Δhep mice exhibited no liver disease at baseline, but developed acute biochemical and histologic liver injury in response to a dietary challenge with fructose for 4 wk. Fructose-mediated liver injury in Xbp1Δhep mice coincided with heightened IRE1α activity, as demonstrated by cJun phosphorylation and regulated IRE1α -dependent RNA decay (RIDD). Activation of eIF2α was also evident, with associated up-regulation of the pro-apoptotic molecules CHOP, BIM and PUMA. To determine whether the adverse consequences of liver-specific XBP1 deletion were due to XBP1 loss or heightened IRE1α activity, we repeated a fructose challenge in mice with liver-specific deletion of both XBP1 and IRE1α (Xbp1Δhep;IRE1αΔhep). Xbp1Δhep;IRE1αΔhep mice were protected from fructose-mediated liver injury and failed to exhibit any of the signs of ER stress seen in mice lacking XBP1 alone. The protective effect of IRE1α deletion persisted even with long-term exposure to fructose. Xbp1Δhep mice developed liver fibrosis at 16 wk, but Xbp1Δhep;IRE1αΔhep mice did not. Overall, the results indicate that the deleterious effects of hepatocyte-specific XBP1 deletion are due primarily to hyperactivation of IRE1α. They support further exploration of IRE1α as a contributor to acute and chronic liver diseases.

scholarly journals A Reproducible Data Analysis Workflow

2021 ◽  
Vol 1 ◽  
Author(s):  
Aaron Peikert ◽  
Andreas M. Brandmaier
Keyword(s):  
Scientific Productivity ◽  
Seamless Integration ◽  
Report Generation ◽  
Version Management ◽  
Computing Platform ◽  
Analysis Workflow ◽  
Cross Platform ◽  
Document Generation ◽  
Statistical Results

In this tutorial, we describe a workflow to ensure long-term reproducibility of R-based data analyses. The workflow leverages established tools and practices from software engineering. It combines the benefits of various open-source software tools including R Markdown, Git, Make, and Docker, whose interplay ensures seamless integration of version management, dynamic report generation conforming to various journal styles, and full cross-platform and long-term computational reproducibility. The workflow ensures meeting the primary goals that 1) the reporting of statistical results is consistent with the actual statistical results (dynamic report generation), 2) the analysis exactly reproduces at a later point in time even if the computing platform or software is changed (computational reproducibility), and 3) changes at any time (during development and post-publication) are tracked, tagged, and documented while earlier versions of both data and code remain accessible. While the research community increasingly recognizes dynamic document generation and version management as tools to ensure reproducibility, we demonstrate with practical examples that these alone are not sufficient to ensure long-term computational reproducibility. Combining containerization, dependence management, version management, and dynamic document generation, the proposed workflow increases scientific productivity by facilitating later reproducibility and reuse of code and data.

scholarly journals Infant functional connectivity fingerprints predict long-term language and pre-literacy outcomes

2020 ◽  
Author(s):  
Xi Yu ◽  
Silvina Ferradal ◽  
Danielle D. Sliva ◽  
Jade Dunstan ◽  
Clarisa Carruthers ◽  
...  
Keyword(s):  
Functional Connectivity ◽  
Cognitive Abilities ◽  
Functional Organization ◽  
Literacy Skills ◽  
Language And Literacy ◽  
Functional Connections ◽  
Literacy Outcomes ◽  
Functional Brain Networks ◽  
Longitudinal Imaging

AbstractFunctional brain networks undergo extensive development within the first few years of life. Previous studies have linked infant functional connectivity to cognitive abilities in toddlerhood. However, little is known regarding the long-term relevance of functional connections established in infancy for the protracted development of higher-order abilities of language and literacy. Employing a five-year longitudinal imaging project starting in infancy, this study utilizes resting-state functional MRI to demonstrate prospective associations between infant functional connectivity fingerprints and subsequent language and foundational literacy skills at a mean age of 6.5. These longitudinal associations are preserved when key environmental influences are controlled for and are independent of emergent language abilities in infancy, suggesting early development of functional network characteristics in supporting the acquisition of high-order language and pre-literacy skills. Altogether, the current results highlight the importance of functional organization established in infancy as a neural scaffold underlying the learning process of complex cognitive functions.

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