Establishment and Characterization of SV40 T-Antigen Immortalized Human Liver Cells

Author(s):  
Masayoshi Namba ◽  
Yoshio Kano ◽  
Li-yan Bai ◽  
Koichiro Mihara ◽  
Masahiro Miyazaki
Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1357
Author(s):  
Niki Alevra Sarika ◽  
Valéry L. Payen ◽  
Maximilien Fléron ◽  
Joachim Ravau ◽  
Davide Brusa ◽  
...  

The lack of robust methods to preserve, purify and in vitro maintain the phenotype of the human liver’s highly specialized parenchymal and non-parenchymal cell types importantly hampers their exploitation for the development of research and clinical applications. There is in this regard a growing interest in the use of tissue-specific extracellular matrix (ECM) to provide cells with an in vitro environment that more closely resembles that of the native tissue. In the present study, we have developed a method that allows for the isolation and downstream application of the human liver’s main cell types from cryopreserved material. We also isolated and solubilized human liver ECM (HL-ECM), analyzed its peptidomic and proteomic composition by mass spectrometry and evaluated its interest for the culture of distinct primary human liver cells. Our analysis of the HL-ECM revealed proteomic diversity, type 1 collagen abundance and partial loss of integrity following solubilization. Solubilized HL-ECM was evaluated either as a coating or as a medium supplement for the culture of human primary hepatocytes, hepatic stellate cells and liver sinusoidal endothelial cells. Whereas the solubilized HL-ECM was suitable for cell culture, its impact on the phenotype and/or functionality of the human liver cells was limited. Our study provides a first detailed characterization of solubilized HL-ECM and a first report of its influence on the culture of distinct human primary liver cells.


Intervirology ◽  
1982 ◽  
Vol 17 (4) ◽  
pp. 222-227 ◽  
Author(s):  
Johannes Buitenwerf ◽  
Wim de Jong ◽  
Ans van Strien ◽  
Jan van der Noordaa

Author(s):  
Reza Afrisham ◽  
Sahar Sadegh-Nejadi ◽  
Reza Meshkani ◽  
Solaleh Emamgholipour ◽  
Molood Bagherieh ◽  
...  

Introduction: Obesity is a disorder with low-grade chronic inflammation that plays a key role in the hepatic inflammation and steatosis. Moreover, there are studies to support the role of exosomes in the cellular communications, the regulation of metabolic homeostasis and immunomodulatory activity. Accordingly, we aimed to evaluate the influence of plasma circulating exosomes derived from females with normal-weight and obesity on the secretion of inflammatory cytokines in human liver cells. Methods: Plasma circulating exosomes were isolated from four normal (N-Exo) and four obese (O-Exo) women. The exosomes were characterized and approved for CD63 expression (common exosomal protein marker) and morphology/size using the western blot and TEM methods, respectively. The exosomes were used for stimulation of HepG2 cells in vitro. After 24 h incubation, the protein levels of TNF-α,IL-6, and IL-1β were measured in the culture supernatant of HepG2 cells using the ELISA kit. Results: The protein levels of IL-6 and TNF-α in the cells treated with O-Exo and N-Exo reduced significantly in comparison with control group (P=0.039 and P<0.001 respectively), while significance differences were not found between normal and obese groups (P=0.808, and P=0.978 respectively). However, no significant differences were found between three groups in term of IL-1β levels (P=0.069). Based on the correlation analysis, the protein levels of IL-6 were positively correlated with TNF-α (r 0.978, P<0.001). Conclusion: These findings suggest that plasma circulating exosomes have probably anti-inflammatory properties independently from body mass index and may decrease the secretion of inflammatory cytokines in liver. However, further investigations in vitro and in vivo are needed to address the anti-inflammatory function of N-Exo and O-Exo in human liver cells and/or other cells.


1995 ◽  
Vol 28 (1) ◽  
pp. 118-128 ◽  
Author(s):  
MARQUÉ D. TODD ◽  
MICHAEL J. LEE ◽  
JULIE L. WILLIAMS ◽  
JOHN M. NALEZNY ◽  
PAULINE GEE ◽  
...  

2010 ◽  
Vol 30 (6) ◽  
pp. 566-573 ◽  
Author(s):  
Saura C. Sahu ◽  
Michael W. O'Donnell ◽  
Paddy L. Wiesenfeld

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