scholarly journals Decellularised Human Liver is too Heterogeneous for Designing a Generic ECM-mimic Hepatic Scaffold

2016 ◽  
Author(s):  
Giorgio Mattei ◽  
Chiara Magliaro ◽  
Andrea Pirone ◽  
Arti Ahluwalia
Keyword(s):  
Human Liver ◽  
Unit Area ◽  
Biochemical Properties ◽  
Matrix Composition ◽  
Hepatic Tissue ◽  
Short Supply ◽  
3 Dimensional ◽  
Ecm Extracellular Matrix ◽  
Donor Variability ◽  
Human Livers

AbstractDecellularised human livers are considered the perfect ECM (extracellular matrix) surrogate because both 3-dimensional architecture and biological features of the hepatic microenvironment are thought to be preserved. However, donor human livers are in chronically short supply, both for transplantation or as decellularised scaffolds, and will become even scarcer as life expectancy increases. It is hence of interest to determine the structural and biochemical properties of human hepatic ECM to derive design criteria for engineering bio-mimetic scaffolds. The intention of this work was to obtain quantitative design specifications for fabricating scaffolds for hepatic tissue engineering using human livers as a template. To this end, hepatic samples from 5 human donors were decellularised using a protocol shown to reproducibly conserve matrix composition and micro-structure in porcine livers. The decellularisation outcome was evaluated through histological and quantitative image analyses to evaluate cell removal, protein and glycosaminoglycan content per unit area. Applying the same decellularisation protocol to human liver samples obtained from 5 different donors yielded 5 different outcomes. Only 1 liver out of 5 was completely decellularised, while the other 4 showed different levels of remaining cells. Moreover, protein and glycosaminoglycan content per unit area after decellularisation were also found to be donor-dependent. The donor-to-donor variability of human livers thus precludes their use as templates for engineering a generic 'one-size fits all' ECM-mimic hepatic scaffold.

scholarly journals Energy determinants GAPDH and NDPK act as genetic modifiers for hepatocyte inclusion formation

2011 ◽  
Vol 195 (2) ◽  
pp. 217-229 ◽  
Author(s):  
Natasha T. Snider ◽  
Sujith V.W. Weerasinghe ◽  
Amika Singla ◽  
Jessica M. Leonard ◽  
Shinichiro Hanada ◽  
...  
Keyword(s):  
Liver Injury ◽  
Human Liver ◽  
Nuclear Translocation ◽  
Nucleoside Diphosphate Kinase ◽  
Isolated Hepatocytes ◽  
Dependent Manner ◽  
Genetic Modifiers ◽  
C3h Mice ◽  
Normal Human ◽  
Human Livers

Genetic factors impact liver injury susceptibility and disease progression. Prominent histological features of some chronic human liver diseases are hepatocyte ballooning and Mallory-Denk bodies. In mice, these features are induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) in a strain-dependent manner, with the C57BL and C3H strains showing high and low susceptibility, respectively. To identify modifiers of DDC-induced liver injury, we compared C57BL and C3H mice using proteomic, biochemical, and cell biological tools. DDC elevated reactive oxygen species (ROS) and oxidative stress enzymes preferentially in C57BL livers and isolated hepatocytes. C57BL livers and hepatocytes also manifested significant down-regulation, aggregation, and nuclear translocation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). GAPDH knockdown depleted bioenergetic and antioxidant enzymes and elevated hepatocyte ROS, whereas GAPDH overexpression decreased hepatocyte ROS. On the other hand, C3H livers had higher expression and activity of the energy-generating nucleoside-diphosphate kinase (NDPK), and knockdown of hepatocyte NDPK augmented DDC-induced ROS formation. Consistent with these findings, cirrhotic, but not normal, human livers contained GAPDH aggregates and NDPK complexes. We propose that GAPDH and NDPK are genetic modifiers of murine DDC-induced liver injury and potentially human liver disease.

scholarly journals Subtoxic Concentrations of Hepatotoxic Drugs Lead to Kupffer Cell Activation in a HumanIn VitroLiver Model: An Approach to Study DILI

2015 ◽  
Vol 2015 ◽  
pp. 1-14 ◽  
Author(s):  
Victoria Kegel ◽  
Elisa Pfeiffer ◽  
Britta Burkhardt ◽  
Jia L. Liu ◽  
Katrin Zeilinger ◽  
...  
Keyword(s):  
Cell Activation ◽  
Cell Activity ◽  
Hepatic Tissue ◽  
Oxygen Intermediates ◽  
Drug Induced ◽  
Xtt Assay ◽  
Liver Model ◽  
Donor Variability ◽  
Stress Damage

Drug induced liver injury (DILI) is an idiosyncratic adverse drug reaction leading to severe liver damage. Kupffer cells (KC) sense hepatic tissue stress/damage and therefore could be a tool for the estimation of consequent effects associated with DILI. Aim of the present study was to establish a humanin vitroliver model for the investigation of immune-mediated signaling in the pathogenesis of DILI. Hepatocytes and KC were isolated from human liver specimens. The isolated KC yield was1.2±0.9×106cells/g liver tissue with a purity of >80%. KC activation was investigated by the measurement of reactive oxygen intermediates (ROI, DCF assay) and cell activity (XTT assay). The initial KC activation levels showed broad donor variability. Additional activation of KC using supernatants of hepatocytes treated with hepatotoxic drugs increased KC activity and led to donor-dependent changes in the formation of ROI compared to KC incubated with supernatants from untreated hepatocytes. Additionally, a compound- and donor-dependent increase in proinflammatory cytokines or in anti-inflammatory cytokines was detected. In conclusion, KC related immune signaling in hepatotoxicity was successfully determined in a newly establishedin vitroliver model. KC were able to detect hepatocyte stress/damage and to transmit a donor- and compound-dependent immune response via cytokine production.

Influence of Preservation Solution on the Isolation and Culture of Human Hepatocytes from Liver Grafts

2005 ◽  
Vol 14 (10) ◽  
pp. 837-843 ◽  
Author(s):  
Alfonso Serralta ◽  
Maria Teresa Donato ◽  
Amparo Martinez ◽  
Eugenia Pareja ◽  
Francisco Orbis ◽  
...  
Keyword(s):  
Cell Viability ◽  
Hepatic Tissue ◽  
Uw Solution ◽  
University Of Wisconsin ◽  
Viable Cells ◽  
Successful Isolation ◽  
Preservation Solutions ◽  
Collagenase Perfusion ◽  
Human Livers

A major problem for the isolation and transplantation of hepatocytes is the lack of resources for obtaining viable hepatocytes. Improving this situation would enhance hepatic cell transplantation programs. Our objective was to evaluate the influence of the preservation solutions used during organ retrieval on the quality of hepatocytes isolated from liver tissue. We compared the results of the collagenase perfusion technique for isolation of hepatocytes in human livers flushed with University of Wisconsin (UW) and Celsior preservation solutions. Yield (number of viable cells per gram of tissue), cellular viability, efficiency of cells to attach to culture plates and form a monolayer, and drug metabolizing competence of the hepatocytes were measured. Successful isolation was achieved in 63% of the procedures using the UW solution and 100% of the procedures using the Celsior solution. In the UW group, significantly lower cell viability (38 ± 41% vs. 79 ± 14%, p < 0.05), yield of cells (4.0 ± 5.2 × 106 vs. 8.2 ± 5.6 × 106 cells/g, p < 0.05), and protein content at 24 h of culture (0.6 ± 0.6 vs. 1.2 ± 0.3 mg protein per plate, p < 0.05) than in Celsior solution were found. However, similar values of P450 activities were found in both groups. The more successful isolation, better yield, and higher cell viability obtained from human liver grafts preserved in Celsior solution, in comparison to UW solution, suggest Celsior solution as the most appropriate for preserving cadaveric hepatic tissue to be used for hepatocyte harvesting.

3D bio-printing ofhuman hepatic tissue using human liver extracellular matrix as tissue-specificbioink

2018 ◽  
Vol 68 ◽  
pp. S55
Author(s):  
L. Frenguelli ◽  
Z. Zhang ◽  
L. Chikh ◽  
W. Al-Akkad ◽  
K. Rombouts ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Human Liver ◽  
Hepatic Tissue

scholarly journals IL-2high tissue-resident T cells in the human liver: Sentinels for hepatotropic infection

2017 ◽  
Vol 214 (6) ◽  
pp. 1567-1580 ◽  
Author(s):  
Laura J. Pallett ◽  
Jessica Davies ◽  
Emily J. Colbeck ◽  
Francis Robertson ◽  
Navjyot Hansi ◽  
...  
Keyword(s):  
T Cells ◽  
Human Liver ◽  
Memory T Cells ◽  
Local Constraints ◽  
Cell Pool ◽  
Mediate Immunity ◽  
B Virus ◽  
Human Livers ◽  
Inhibitory Molecules ◽  
Resident Memory T Cells

The liver provides a tolerogenic immune niche exploited by several highly prevalent pathogens as well as by primary and metastatic tumors. We have sampled healthy and hepatitis B virus (HBV)–infected human livers to probe for a subset of T cells specialized to overcome local constraints and mediate immunity. We characterize a population of T-betloEomesloBlimp-1hiHobitlo T cells found within the intrahepatic but not the circulating memory CD8 T cell pool expressing liver-homing/retention markers (CD69+CD103+ CXCR6+CXCR3+). These tissue-resident memory T cells (TRM) are preferentially expanded in patients with partial immune control of HBV infection and can remain in the liver after the resolution of infection, including compartmentalized responses against epitopes within all major HBV proteins. Sequential IL-15 or antigen exposure followed by TGFβ induces liver-adapted TRM, including their signature high expression of exhaustion markers PD-1 and CD39. We suggest that these inhibitory molecules, together with paradoxically robust, rapid, cell-autonomous IL-2 and IFNγ production, equip liver CD8 TRM to survive while exerting local noncytolytic hepatic immunosurveillance.

Hepatic Tissue Engineering on 3-Dimensional Biodegradable Polymers within a Pulsatile Flow Bioreactor

2001 ◽  
Vol 18 (3) ◽  
pp. 196-203 ◽  
Author(s):  
É. Török ◽  
J.-M. Pollok ◽  
P.X. Ma ◽  
C. Vogel ◽  
M. Dandri ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Biodegradable Polymers ◽  
Pulsatile Flow ◽  
Hepatic Tissue ◽  
3 Dimensional

scholarly journals Extracellular Matrix Optimization for Enhanced Physiological Relevance in Hepatic Tissue-Chips

Polymers ◽  
2021 ◽  
Vol 13 (17) ◽  
pp. 3016
Author(s):  
Abdul Rahim Chethikkattuveli Salih ◽  
Kinam Hyun ◽  
Arun Asif ◽  
Afaque Manzoor Soomro ◽  
Hafiz Muhammad Umer Farooqi ◽  
...  
Keyword(s):  
Image Processing ◽  
Extracellular Matrix ◽  
Drug Screening ◽  
Human Liver ◽  
Growth Patterns ◽  
Surface Coatings ◽  
Hepatic Tissue ◽  
Tissue Formation ◽  
Tissue Development

The cellular microenvironment is influenced explicitly by the extracellular matrix (ECM), the main tissue support biomaterial, as a decisive factor for tissue growth patterns. The recent emergence of hepatic microphysiological systems (MPS) provide the basic physiological emulation of the human liver for drug screening. However, engineering microfluidic devices with standardized surface coatings of ECM may improve MPS-based organ-specific emulation for improved drug screening. The influence of surface coatings of different ECM types on tissue development needs to be optimized. Additionally, an intensity-based image processing tool and transepithelial electrical resistance (TEER) sensor may assist in the analysis of tissue formation capacity under the influence of different ECM types. The current study highlights the role of ECM coatings for improved tissue formation, implying the additional role of image processing and TEER sensors. We studied hepatic tissue formation under the influence of multiple concentrations of Matrigel, collagen, fibronectin, and poly-L-lysine. Based on experimental data, a mathematical model was developed, and ECM concentrations were validated for better tissue development. TEER sensor and image processing data were used to evaluate the development of a hepatic MPS for human liver physiology modeling. Image analysis data for tissue formation was further strengthened by metabolic quantification of albumin, urea, and cytochrome P450. Standardized ECM type for MPS may improve clinical relevance for modeling hepatic tissue microenvironment, and image processing possibly enhance the tissue analysis of the MPS.

scholarly journals Identification of the human liver cytochrome P-450 responsible for coumarin 7-hydroxylase activity

1990 ◽  
Vol 267 (2) ◽  
pp. 365-371 ◽  
Author(s):  
J S Miles ◽  
A W McLaren ◽  
L M Forrester ◽  
M J Glancey ◽  
M A Lang ◽  
...  
Keyword(s):  
Human Liver ◽  
Liver Microsomes ◽  
Microsomal Protein ◽  
Hydroxylase Activity ◽  
Liver Cytochrome ◽  
Partial Length ◽  
Coumarin 7 ◽  
Human Livers ◽  
Cell Expression ◽  
Cytochrome P 450

1. We have constructed a full-length human liver cytochrome P450IIA cDNA from a partial-length clone by oligonucleotide-directed mutagenesis, and subcloned it into the monkey kidney (COS-7) cell expression vector, pSVL. 2. The cDNA encodes a 49 kDa protein with coumarin 7-hydroxylase (COH) activity which cross-reacts with antisera to the mouse cytochrome P-450 isoenzyme responsible for COH activity and comigrates with a human liver microsomal protein. 3. Western blot analysis of a panel of human livers indicates that the level of the 49 kDa protein, detected using antisera to either the mouse COH P-450 or rat P450IIA1 protein, correlates very highly with COH activity. 4. Antisera to the rat P450IIA1 protein can inhibit COH activity in human liver microsomes. Taken together, these data indicate that a member of the P450IIA subfamily is responsible for most, if not all, of the COH activity in human liver.

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