Construction of mouse liver aging model by hepatocyte organoid
Objective Construction of mouse liver senescence model induced rapidly in vitro by 3D organoid culture. Methods Mouse hepatocytes were collected by two-step perfusion method and cultured in 3D. Organoids were preliminarily identified by morphology, qPCR and immunofluorescence. The function of hepato...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2023-04-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-4-554.pdf |
| _version_ | 1797366372603265024 |
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| author | LU Yan, ZHOU Yabo, CHEN Jie |
| author_facet | LU Yan, ZHOU Yabo, CHEN Jie |
| author_sort | LU Yan, ZHOU Yabo, CHEN Jie |
| collection | DOAJ |
| description | Objective Construction of mouse liver senescence model induced rapidly in vitro by 3D organoid culture. Methods Mouse hepatocytes were collected by two-step perfusion method and cultured in 3D. Organoids were preliminarily identified by morphology, qPCR and immunofluorescence. The function of hepatocyte organoid was evaluated by their uptake of LDL. The hepatocyte organoids were treated with oleic acid, and their clonal growth, senescence related secretory phenotype, LDL uptake and ROS level of liver cells were detected. The hepatocytes were further treated with N-acetylcysteine (NAC) to detect the above phenotypic and functional changes. Results A large number of active primary mouse hepatocytes were obtained by two-step perfusion method. The hepatocyte organoid with self-reproduction ability was collected by 3D culture in the matrix gel. The expression of liver marker genes in hepatocyte organoid and primary hepatocytes was shown by qPCR detection. The immunofluorescence results showed that it highly expressed ALB protein. Moreover, hepatocyte organoid had normal uptake function of LDL. After treatment with oleic acid, the growth of hepatocyte organoid was slowed down, the expression of senescence related secretory phenotype genes was up-regulated(P＜0.001), the uptake of LDL was reduced(P＜0.001), and ROS was significantly increased(P＜0.001). The above phenotypes were alleviated by NAC treatment(P＜0.01 or P＜0.001). Conclusions Oleic acid treatment can successfully construct the aging model of mouse liver in vitro. Adding NAC can clear ROS and alleviate the occurrence of aging. |
| first_indexed | 2024-03-08T17:03:13Z |
| format | Article |
| id | doaj.art-a25ac52081074d10ba67d91f1da5cfe6 |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T17:03:13Z |
| publishDate | 2023-04-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-a25ac52081074d10ba67d91f1da5cfe62024-01-04T07:32:42ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252023-04-0143455455910.16352/j.issn.1001-6325.2023.04.0554Construction of mouse liver aging model by hepatocyte organoidLU Yan, ZHOU Yabo, CHEN Jie0National Key Laboratory of Medical Molecular Biology, Department of Immunology, Institute of Basic Medical Sciences CAMS, School of Basic Medicine PUMC, Beijing 100005,ChinaObjective Construction of mouse liver senescence model induced rapidly in vitro by 3D organoid culture. Methods Mouse hepatocytes were collected by two-step perfusion method and cultured in 3D. Organoids were preliminarily identified by morphology, qPCR and immunofluorescence. The function of hepatocyte organoid was evaluated by their uptake of LDL. The hepatocyte organoids were treated with oleic acid, and their clonal growth, senescence related secretory phenotype, LDL uptake and ROS level of liver cells were detected. The hepatocytes were further treated with N-acetylcysteine (NAC) to detect the above phenotypic and functional changes. Results A large number of active primary mouse hepatocytes were obtained by two-step perfusion method. The hepatocyte organoid with self-reproduction ability was collected by 3D culture in the matrix gel. The expression of liver marker genes in hepatocyte organoid and primary hepatocytes was shown by qPCR detection. The immunofluorescence results showed that it highly expressed ALB protein. Moreover, hepatocyte organoid had normal uptake function of LDL. After treatment with oleic acid, the growth of hepatocyte organoid was slowed down, the expression of senescence related secretory phenotype genes was up-regulated(P＜0.001), the uptake of LDL was reduced(P＜0.001), and ROS was significantly increased(P＜0.001). The above phenotypes were alleviated by NAC treatment(P＜0.01 or P＜0.001). Conclusions Oleic acid treatment can successfully construct the aging model of mouse liver in vitro. Adding NAC can clear ROS and alleviate the occurrence of aging.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-4-554.pdfprimary hepatocyte|organoid|oleic acid|reactive oxygen species|aging |
| spellingShingle | LU Yan, ZHOU Yabo, CHEN Jie Construction of mouse liver aging model by hepatocyte organoid Jichu yixue yu linchuang primary hepatocyte|organoid|oleic acid|reactive oxygen species|aging |
| title | Construction of mouse liver aging model by hepatocyte organoid |
| title_full | Construction of mouse liver aging model by hepatocyte organoid |
| title_fullStr | Construction of mouse liver aging model by hepatocyte organoid |
| title_full_unstemmed | Construction of mouse liver aging model by hepatocyte organoid |
| title_short | Construction of mouse liver aging model by hepatocyte organoid |
| title_sort | construction of mouse liver aging model by hepatocyte organoid |
| topic | primary hepatocyte|organoid|oleic acid|reactive oxygen species|aging |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2023-43-4-554.pdf |
| work_keys_str_mv | AT luyanzhouyabochenjie constructionofmouseliveragingmodelbyhepatocyteorganoid |