Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system
Abstract Liver diseases, including NAFLD, are a growing worldwide health concern. Currently, there is a lack of suitable in vitro models that sustain basic primary human hepatocyte (PHH) morphology and functionality while supporting presentation of disease-associated phenotypic characteristics such...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-03-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-57463-7 |
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| author | Justin J. Odanga Sharon M. Anderson Erick K. Breathwaite Sharon C. Presnell Edward L. LeCluyse Jingsong Chen Jessica R. Weaver |
| author_facet | Justin J. Odanga Sharon M. Anderson Erick K. Breathwaite Sharon C. Presnell Edward L. LeCluyse Jingsong Chen Jessica R. Weaver |
| author_sort | Justin J. Odanga |
| collection | DOAJ |
| description | Abstract Liver diseases, including NAFLD, are a growing worldwide health concern. Currently, there is a lack of suitable in vitro models that sustain basic primary human hepatocyte (PHH) morphology and functionality while supporting presentation of disease-associated phenotypic characteristics such as lipid accumulation and inflammasome activation. In TruVivo, an all-human triculture system (hTCS), basic metabolic functions were characterized in PHHs isolated from normal or diseased livers during two-weeks of culture. Decreases in albumin and urea levels and CYP3A4 activity were seen in diseased-origin PHHs compared to normal PHHs along with higher CYP2E1 expression. Positive expression of the macrophage markers CD68 and CD163 were seen in the diseased PHH preparations. Elevated levels of the pro-inflammatory cytokines IL-6 and MCP-1 and the fibrotic markers CK-18 and TGF-β were also measured. Gene expression of FASN, PCK1, and G6PC in the diseased PHHs was decreased compared to the normal PHHs. Further characterization revealed differences in lipogenesis and accumulation of intracellular lipids in normal and diseased PHHs when cultured with oleic acid and high glucose. TruVivo represents a promising new platform to study lipogenic mechanisms in normal and diseased populations due to the preservation of phenotypic differences over a prolonged culture period. |
| first_indexed | 2024-04-24T19:56:51Z |
| format | Article |
| id | doaj.art-3a0a4cf5b93f4ff7800f377f929071e4 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-24T19:56:51Z |
| publishDate | 2024-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-3a0a4cf5b93f4ff7800f377f929071e42024-03-24T12:18:21ZengNature PortfolioScientific Reports2045-23222024-03-0114111310.1038/s41598-024-57463-7Characterization of diseased primary human hepatocytes in an all-human cell-based triculture systemJustin J. Odanga0Sharon M. Anderson1Erick K. Breathwaite2Sharon C. Presnell3Edward L. LeCluyse4Jingsong Chen5Jessica R. Weaver6Institute of Regenerative Med., LifeNet HealthInstitute of Regenerative Med., LifeNet HealthInstitute of Regenerative Med., LifeNet HealthInstitute of Regenerative Med., LifeNet HealthResearch and Development, LifeNet Health LifeSciencesInstitute of Regenerative Med., LifeNet HealthInstitute of Regenerative Med., LifeNet HealthAbstract Liver diseases, including NAFLD, are a growing worldwide health concern. Currently, there is a lack of suitable in vitro models that sustain basic primary human hepatocyte (PHH) morphology and functionality while supporting presentation of disease-associated phenotypic characteristics such as lipid accumulation and inflammasome activation. In TruVivo, an all-human triculture system (hTCS), basic metabolic functions were characterized in PHHs isolated from normal or diseased livers during two-weeks of culture. Decreases in albumin and urea levels and CYP3A4 activity were seen in diseased-origin PHHs compared to normal PHHs along with higher CYP2E1 expression. Positive expression of the macrophage markers CD68 and CD163 were seen in the diseased PHH preparations. Elevated levels of the pro-inflammatory cytokines IL-6 and MCP-1 and the fibrotic markers CK-18 and TGF-β were also measured. Gene expression of FASN, PCK1, and G6PC in the diseased PHHs was decreased compared to the normal PHHs. Further characterization revealed differences in lipogenesis and accumulation of intracellular lipids in normal and diseased PHHs when cultured with oleic acid and high glucose. TruVivo represents a promising new platform to study lipogenic mechanisms in normal and diseased populations due to the preservation of phenotypic differences over a prolonged culture period.https://doi.org/10.1038/s41598-024-57463-7LiverHepatocytesHumanDiseaseNASH |
| spellingShingle | Justin J. Odanga Sharon M. Anderson Erick K. Breathwaite Sharon C. Presnell Edward L. LeCluyse Jingsong Chen Jessica R. Weaver Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system Scientific Reports Liver Hepatocytes Human Disease NASH |
| title | Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system |
| title_full | Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system |
| title_fullStr | Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system |
| title_full_unstemmed | Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system |
| title_short | Characterization of diseased primary human hepatocytes in an all-human cell-based triculture system |
| title_sort | characterization of diseased primary human hepatocytes in an all human cell based triculture system |
| topic | Liver Hepatocytes Human Disease NASH |
| url | https://doi.org/10.1038/s41598-024-57463-7 |
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