A practice-changing culture method relying on shaking substantially increases mitochondrial energy metabolism and functionality of human liver cell lines
Publication Date
April 19, 2018
Authors
Aziza A. A. Adam, Vincent A. Van Der Mark, Joanne M. Donkers, Manon E. Wildenberg, et al
Volume
13
Issue
4
Pages
e0193664
DOI
https://dx.plos.org/10.1371/journal.pone.0193664
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0193664
Scopus
85045943532
Mendeley
http://www.mendeley.com/research/practicechanging-culture-method-relying-shaking-substantially-increases-mitochondrial-energy-metabol
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Mendeley | Further Information

{"title"=>"A practice-changing culture method relying on shaking substantially increases mitochondrial energy metabolism and functionality of human liver cell lines", "type"=>"journal", "authors"=>[{"first_name"=>"Aziza A. A.", "last_name"=>"Adam"}, {"first_name"=>"Vincent A.", "last_name"=>"van der Mark"}, {"first_name"=>"Joanne M.", "last_name"=>"Donkers"}, {"first_name"=>"Manon E.", "last_name"=>"Wildenberg"}, {"first_name"=>"Ronald P. J.", "last_name"=>"Oude Elferink"}, {"first_name"=>"Robert A. F. M.", "last_name"=>"Chamuleau"}, {"first_name"=>"Ruurdtje", "last_name"=>"Hoekstra"}], "year"=>2018, "source"=>"PLOS ONE", "identifiers"=>{"pui"=>"621762328", "pmid"=>"29672606", "doi"=>"10.1371/journal.pone.0193664", "issn"=>"1932-6203", "scopus"=>"2-s2.0-85045943532", "sgr"=>"85045943532"}, "id"=>"c54488c1-80e4-3700-99fe-91f385814c89", "abstract"=>"Practice-changing culturing techniques of hepatocytes are highly required to increase their differentiation. Previously, we found that human liver cell lines HepaRG and C3A acquire higher functionality and increased mitochondrial biogenesis when cultured in the AMC-Bioartificial liver (BAL). Dynamic medium flow (DMF) is one of the major contributors to this stimulatory effect. Recently, we found that DMF-culturing by shaking of HepaRG monolayers resulted in higher mitochondrial biogenesis. Here we further investigated the effect of DMF-culturing on energy metabolism and hepatic functionality of HepaRG and C3A monolayers. HepaRG and C3A DMF-monolayers were incubated with orbital shaking at 60 rpm during the differentiation phase, while control monolayers were maintained statically. Subsequently, energy metabolism and hepatic functionality were compared between static and DMF-cultures. DMF-culturing of HepaRG cells substantially increased hepatic differentiation; transcript levels of hepatic structural genes and hepatic transcription regulators were increased up to 15-fold (Cytochrome P450 3A4) and nuclear translocation of hepatic transcription factor CEBPα was stimulated. Accordingly, hepatic functions were positively affected, including ammonia elimination, urea production, bile acid production, and CYP3A4 activity. DMF-culturing shifted energy metabolism from aerobic glycolysis towards oxidative phosphorylation, as indicated by a decline in lactate production and glucose consumption, and an increase in oxygen consumption. Similarly, DMF-culturing increased mitochondrial energy metabolism and hepatic functionality of C3A cells. In conclusion, simple shaking of monolayer cultures substantially improves mitochondrial energy metabolism and hepatic differentiation of human liver cell lines. This practice-changing culture method may prove to prolong the in-vitro maintenance of primary hepatocytes and increase hepatic differentiation of stem cells.", "link"=>"http://www.mendeley.com/research/practicechanging-culture-method-relying-shaking-substantially-increases-mitochondrial-energy-metabol", "reader_count"=>2, "reader_count_by_academic_status"=>{"Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>1, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Chemistry"=>{"Chemistry"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "group_count"=>0}

Scopus | Further Information

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