An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation

10.3791/61812 ◽  
2021 ◽  
Author(s):  
Mingxiao Feng ◽  
Sara Divall ◽  
Sheng Wu
Keyword(s):  
Primary Hepatocyte ◽  
Hepatocyte Isolation

scholarly journals Marine Fish Primary Hepatocyte Isolation and Culture: New Insights to Enzymatic Dissociation Pancreatin Digestion

2021 ◽  
Vol 18 (4) ◽  
pp. 1380
Author(s):  
Neusa Figueiredo ◽  
Beatriz Matos ◽  
Mário Diniz ◽  
Vasco Branco ◽  
Marta Martins
Keyword(s):  
Cell Viability ◽  
Cell Cultures ◽  
Marine Fish ◽  
Sparus Aurata ◽  
Cost Effective ◽  
Primary Hepatocyte ◽  
Digestion Method ◽  
Trypan Blue Exclusion ◽  
And Function

Primary cell cultures from wild organisms have been gaining relevance in ecotoxicology as they are considered more sensitive than immortalized cell lines and retain the biochemical pathways found in vivo. In this study, the efficacy of two methods for primary hepatocyte cell isolation was compared using liver from two marine fish (Sparus aurata and Psetta maxima): (i) two-step collagenase perfusion and (ii) pancreatin digestion with modifications. Cell cultures were incubated in L-15 medium at 17 ± 1 °C and monitored for up to six days for cell viability and function using the trypan blue exclusion test, MTT test, lactate dehydrogenase (LDH) activity, and ethoxyresorufin O-deethylase (EROD) activity after Benzo[a]Pyrene exposure. The results showed significant differences between the number of viable cells (p < 0.05), the highest number being obtained for the pancreatin digestion method (average = 4.5 ± 1.9 × 107 cells). Moreover, the hepatocytes showed solid adherence to the culture plate and the rounded shape, changing into a triangular/polygonal shape. The cell viability and function obtained by pancreatin digestion were maintained for five days, and the EROD induction after exposure to the B[a]P showed that cells were metabolically active. This study shows that the optimized pancreatin digestion method is a valid, cost-effective, and simple alternative to the standard perfusion method for the isolation of primary hepatocytes from fish and is suitable for ecotoxicological studies involving marine pollutants, such as PAHs.

Assessing Mechanisms of Ketoconazole and Amiodarone Induced Steatohepatitis in an Organotypic Human Primary Hepatocyte Liver Model

2016 ◽  
Vol 64 (2) ◽  
pp. S226-S227
Author(s):  
R. Figler ◽  
S. Marukian ◽  
M. Collado ◽  
D. Manka ◽  
M. Lawson ◽  
...  
Keyword(s):  
Primary Hepatocyte ◽  
Liver Model

scholarly journals Substrate stiffness regulates primary hepatocyte functions

RSC Advances ◽  
2015 ◽  
Vol 5 (99) ◽  
pp. 80956-80966 ◽  
Author(s):  
Vaishaali Natarajan ◽  
Eric J. Berglund ◽  
Dorothy X. Chen ◽  
Srivatsan Kidambi
Keyword(s):  
Liver Fibrosis ◽  
Alcohol Abuse ◽  
Metabolic Disorders ◽  
Viral Infections ◽  
Substrate Stiffness ◽  
Primary Hepatocyte ◽  
Chronic Injuries

Liver fibrosis occurs as a consequence of chronic injuries from viral infections, metabolic disorders, and alcohol abuse.

Primary hepatocyte survival on non-integrin-recognizable matrices without the activation of Akt signaling

Biomaterials ◽  
2007 ◽  
Vol 28 (6) ◽  
pp. 1093-1104 ◽  
Author(s):  
Takashi Hoshiba ◽  
Hikaru Nagahara ◽  
Chong-Su Cho ◽  
Yoh-ichi Tagawa ◽  
Toshihiro Akaike
Keyword(s):  
Akt Signaling ◽  
Primary Hepatocyte

scholarly journals A Rat Primary Hepatocyte Culture Model for Aging Studies

2008 ◽  
Vol 37 (1) ◽  
Author(s):  
Swapna V. Shenvi ◽  
Brian M. Dixon ◽  
Kate Petersen Shay ◽  
Tory M. Hagen
Keyword(s):  
Hepatocyte Culture ◽  
Primary Hepatocyte ◽  
Primary Hepatocyte Culture ◽  
Culture Model ◽  
Aging Studies

Bulk Droplet Vitrification for Primary Hepatocyte Preservation

10.3791/60250 ◽  
2019 ◽  
Author(s):  
Reinier J. de Vries ◽  
Peony D. Banik ◽  
Sonal Nagpal ◽  
Lindong Weng ◽  
Sinan Ozer ◽  
...  
Keyword(s):  
Primary Hepatocyte ◽  
Droplet Vitrification
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