Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells
Objective: to obtain long-lived proliferating cells with progenitor features by dedifferentiation of mature rat hepatocytes using combinations of small molecules.Materials and Methods. Hepatocytes isolated from rat liver by perfusion were cultured in the presence of a cocktail of three small molecul...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | Russian |
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Federal Research Center of Transplantology and Artificial Organs named after V.I.Shumakov
2021-09-01
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| Series: | Vestnik Transplantologii i Iskusstvennyh Organov |
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| Online Access: | https://journal.transpl.ru/vtio/article/view/1383 |
| _version_ | 1797871946854039552 |
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| author | A. M. Grigoriev I. V. Kholodenko A. Y. Lupatov R. V. Kholodenko L. A. Kirsanova Y. B. Basok K. N. Yarygin V. I. Sevastianov |
| author_facet | A. M. Grigoriev I. V. Kholodenko A. Y. Lupatov R. V. Kholodenko L. A. Kirsanova Y. B. Basok K. N. Yarygin V. I. Sevastianov |
| author_sort | A. M. Grigoriev |
| collection | DOAJ |
| description | Objective: to obtain long-lived proliferating cells with progenitor features by dedifferentiation of mature rat hepatocytes using combinations of small molecules.Materials and Methods. Hepatocytes isolated from rat liver by perfusion were cultured in the presence of a cocktail of three small molecules – Wnt signaling pathway activator (CHIR99021), TGF-β inhibitors (A83-01) and ROCK kinase (Y27632). The morphological characteristics and growth features of the culture were assessed using fluorescence and phase-contrast microscopy during cell culture. Cell proliferative activity was analyzed using real-time time-lapse imaging. The expression of surface and intracellular markers was analyzed using flow cytometry and high-resolution fluorescence microscopy.Results. Using a cocktail of small molecules, Y-27632, A-83-01, and CHIR99021, long-lived proliferating cells that express progenitor cell markers, such as α-fetoprotein and HNF4α, were obtained from mature rat hepatocytes. The cells had hepatocyte-like morphology and formed discrete clusters of proliferating cells, forming a single cell layer during culturing. Removal of the small molecules from the medium led to expansion of fibroblast-like cells and elimination of potentially progenitor hepatocyte-like cells.Conclusion. Proliferating progenitor cells can be obtained by dedifferentiation of mature hepatocytes. |
| first_indexed | 2024-04-10T00:50:57Z |
| format | Article |
| id | doaj.art-a31f156a344e46008e2ed560f9dc291e |
| institution | Directory Open Access Journal |
| issn | 1995-1191 |
| language | Russian |
| last_indexed | 2024-04-10T00:50:57Z |
| publishDate | 2021-09-01 |
| publisher | Federal Research Center of Transplantology and Artificial Organs named after V.I.Shumakov |
| record_format | Article |
| series | Vestnik Transplantologii i Iskusstvennyh Organov |
| spelling | doaj.art-a31f156a344e46008e2ed560f9dc291e2023-03-13T10:37:28ZrusFederal Research Center of Transplantology and Artificial Organs named after V.I.ShumakovVestnik Transplantologii i Iskusstvennyh Organov1995-11912021-09-0123314816110.15825/1995-1191-2021-3-148-1611034Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cellsA. M. Grigoriev0I. V. Kholodenko1A. Y. Lupatov2R. V. Kholodenko3L. A. Kirsanova4Y. B. Basok5K. N. Yarygin6V. I. Sevastianov7Shumakov National Medical Research Center of Transplantology and Artificial OrgansOrekhovich Institute of Biomedical ChemistryOrekhovich Institute of Biomedical ChemistryShemyakin and Ovchinnikov Institute of Bioorganic ChemistryShumakov National Medical Research Center of Transplantology and Artificial OrgansShumakov National Medical Research Center of Transplantology and Artificial OrgansOrekhovich Institute of Biomedical ChemistryShumakov National Medical Research Center of Transplantology and Artificial OrgansObjective: to obtain long-lived proliferating cells with progenitor features by dedifferentiation of mature rat hepatocytes using combinations of small molecules.Materials and Methods. Hepatocytes isolated from rat liver by perfusion were cultured in the presence of a cocktail of three small molecules – Wnt signaling pathway activator (CHIR99021), TGF-β inhibitors (A83-01) and ROCK kinase (Y27632). The morphological characteristics and growth features of the culture were assessed using fluorescence and phase-contrast microscopy during cell culture. Cell proliferative activity was analyzed using real-time time-lapse imaging. The expression of surface and intracellular markers was analyzed using flow cytometry and high-resolution fluorescence microscopy.Results. Using a cocktail of small molecules, Y-27632, A-83-01, and CHIR99021, long-lived proliferating cells that express progenitor cell markers, such as α-fetoprotein and HNF4α, were obtained from mature rat hepatocytes. The cells had hepatocyte-like morphology and formed discrete clusters of proliferating cells, forming a single cell layer during culturing. Removal of the small molecules from the medium led to expansion of fibroblast-like cells and elimination of potentially progenitor hepatocyte-like cells.Conclusion. Proliferating progenitor cells can be obtained by dedifferentiation of mature hepatocytes.https://journal.transpl.ru/vtio/article/view/1383hepatocytesdedifferentiationhepatic progenitor cellssmall moleculessignaling pathwaysy-27632a-83-01chir99021 |
| spellingShingle | A. M. Grigoriev I. V. Kholodenko A. Y. Lupatov R. V. Kholodenko L. A. Kirsanova Y. B. Basok K. N. Yarygin V. I. Sevastianov Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells Vestnik Transplantologii i Iskusstvennyh Organov hepatocytes dedifferentiation hepatic progenitor cells small molecules signaling pathways y-27632 a-83-01 chir99021 |
| title | Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| title_full | Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| title_fullStr | Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| title_full_unstemmed | Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| title_short | Mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| title_sort | mature rat hepatocyte dedifferentiation into long lived proliferating hepatic progenitor cells |
| topic | hepatocytes dedifferentiation hepatic progenitor cells small molecules signaling pathways y-27632 a-83-01 chir99021 |
| url | https://journal.transpl.ru/vtio/article/view/1383 |
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