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Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome

AIM: To develop a novel hepatocyte serum-free medium based on sericin, and to explore the effect of sericin on the hepatocyte transcriptome. METHODS: A controlled trial comparing novel serum-free medium and other media: C3A cells were cultured in our novel serum-free medium, HepatoZYME, complete med...

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Autores principales: Huang, Yun, Peng, Qing, Li, Hai-Yan, Jia, Zhi-Dong, Li, Yang, Gao, Yi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Baishideng Publishing Group Inc 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6092578/
https://www.ncbi.nlm.nih.gov/pubmed/30122879
http://dx.doi.org/10.3748/wjg.v24.i30.3398
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author Huang, Yun
Peng, Qing
Li, Hai-Yan
Jia, Zhi-Dong
Li, Yang
Gao, Yi
author_facet Huang, Yun
Peng, Qing
Li, Hai-Yan
Jia, Zhi-Dong
Li, Yang
Gao, Yi
author_sort Huang, Yun
collection PubMed
description AIM: To develop a novel hepatocyte serum-free medium based on sericin, and to explore the effect of sericin on the hepatocyte transcriptome. METHODS: A controlled trial comparing novel serum-free medium and other media: C3A cells were cultured in our novel serum-free medium, HepatoZYME, complete medium (DMEM/F12 with 100 mL/L FBS), and DMEM/F12, and then cell attachment, proliferation, and function as well as the biocompatibility of the media were assessed. A comparative study of serum-free media with or without 2 mg/mL sericin: the effect of sericin on C3A growth was assessed by cell viability and proliferation, the effect of sericin on C3A cell cycle distribution was determined by flow cytometry, and the effect of sericin on the C3A transcriptome was assessed by gene-chip array and RT-qPCR. RESULTS: More C3A cells attached to the plate containing our serum-free medium than to those containing HepatoZYME and DMEM/F12 at 24 h post-seeding. Both the viability and proliferation rate of C3A cells in sericin-based serum-free medium were superior to those of cells in HepatoZYME and DMEM/F12 (P < 0.001). The content of albumin and urea in our serum-free medium was significantly higher than that in HepatoZYME and DMEM/F12 throughout the whole culture period (P < 0.001) and was similar to that in complete medium at day 3, 4, and 5. In part 2, cell viability and proliferation were greater in the presence of 2 mg/mL sericin (P < 0.001), as was the proportion of cells in S phase (16.21% ± 0.98% vs 12.61% ± 0.90%, P < 0.01). Gene-chip array analysis indicated that the expression of CCR6, EGFR, and FOS were up-regulated by 2 mg/mL sericin, and RT-qPCR revealed that the expression of CCR6, EGFR, FOS, AKT1, JNK1, NFkB1, MMP-9, MEK2, ERK1/2 and MYC was up-regulated by 2 mg/mL sericin (P < 0.05). CONCLUSION: We developed a novel hepatocyte serum-free medium. Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-κB pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway.
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spelling pubmed-60925782018-08-17 Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome Huang, Yun Peng, Qing Li, Hai-Yan Jia, Zhi-Dong Li, Yang Gao, Yi World J Gastroenterol Basic Study AIM: To develop a novel hepatocyte serum-free medium based on sericin, and to explore the effect of sericin on the hepatocyte transcriptome. METHODS: A controlled trial comparing novel serum-free medium and other media: C3A cells were cultured in our novel serum-free medium, HepatoZYME, complete medium (DMEM/F12 with 100 mL/L FBS), and DMEM/F12, and then cell attachment, proliferation, and function as well as the biocompatibility of the media were assessed. A comparative study of serum-free media with or without 2 mg/mL sericin: the effect of sericin on C3A growth was assessed by cell viability and proliferation, the effect of sericin on C3A cell cycle distribution was determined by flow cytometry, and the effect of sericin on the C3A transcriptome was assessed by gene-chip array and RT-qPCR. RESULTS: More C3A cells attached to the plate containing our serum-free medium than to those containing HepatoZYME and DMEM/F12 at 24 h post-seeding. Both the viability and proliferation rate of C3A cells in sericin-based serum-free medium were superior to those of cells in HepatoZYME and DMEM/F12 (P < 0.001). The content of albumin and urea in our serum-free medium was significantly higher than that in HepatoZYME and DMEM/F12 throughout the whole culture period (P < 0.001) and was similar to that in complete medium at day 3, 4, and 5. In part 2, cell viability and proliferation were greater in the presence of 2 mg/mL sericin (P < 0.001), as was the proportion of cells in S phase (16.21% ± 0.98% vs 12.61% ± 0.90%, P < 0.01). Gene-chip array analysis indicated that the expression of CCR6, EGFR, and FOS were up-regulated by 2 mg/mL sericin, and RT-qPCR revealed that the expression of CCR6, EGFR, FOS, AKT1, JNK1, NFkB1, MMP-9, MEK2, ERK1/2 and MYC was up-regulated by 2 mg/mL sericin (P < 0.05). CONCLUSION: We developed a novel hepatocyte serum-free medium. Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-κB pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway. Baishideng Publishing Group Inc 2018-08-14 2018-08-14 /pmc/articles/PMC6092578/ /pubmed/30122879 http://dx.doi.org/10.3748/wjg.v24.i30.3398 Text en ©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved. http://creativecommons.org/licenses/by-nc/4.0/ This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial.
spellingShingle Basic Study
Huang, Yun
Peng, Qing
Li, Hai-Yan
Jia, Zhi-Dong
Li, Yang
Gao, Yi
Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title_full Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title_fullStr Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title_full_unstemmed Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title_short Novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
title_sort novel sericin-based hepatocyte serum-free medium and sericin’s effect on hepatocyte transcriptome
topic Basic Study
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6092578/
https://www.ncbi.nlm.nih.gov/pubmed/30122879
http://dx.doi.org/10.3748/wjg.v24.i30.3398
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