Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture
Rat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in...
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2024-12-01
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| author | Shanshan Zheng Masato Nakagawa Yanan Gong Yasuhiko Matsushima Satoshi Sasayama Shunsuke Baba Yoshitomo Honda |
| author_facet | Shanshan Zheng Masato Nakagawa Yanan Gong Yasuhiko Matsushima Satoshi Sasayama Shunsuke Baba Yoshitomo Honda |
| author_sort | Shanshan Zheng |
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| description | Rat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in DPSCs after 3–26 passages (3P–26P). The results show that cell morphology and nuclear size increase proportionally with passage number. The phosphorylated histone H2A.X (γ-H2A.X) positive cells (indicating DNA damage) increased significantly earlier than the 4-Hydroxynonenal (4-HNE) stained cells (indicating an abundance of intracellular reactive oxygen species). Compared to the cells subjected to 3P and 5P, the cells subjected to 15P showed reduced proliferation despite being positive for Ki67. Furthermore, cell growth was completely arrested after 26P. The senescence markers, senescence-associated β-galactosidase (SA-β-gal) and p16, exhibited similar expression patterns that were not correlated with those of p21 and urokinase-type plasminogen activator receptor (uPAR). Nearly all cells expressed SA-β-gal and p16 after 26P, whereas only half expressed p21 and uPAR. These results will contribute to understanding the characteristics of DPSCs toward replicative senescence, which are applicable to elucidate mechanisms related to regenerative medicine and stem cell aging. |
| format | Article |
| id | doaj-art-307b88ae44c04ec0bdff8da2f0db1a22 |
| institution | DOAJ |
| issn | 2076-3417 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
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| spelling | doaj-art-307b88ae44c04ec0bdff8da2f0db1a222025-08-20T02:50:18ZengMDPI AGApplied Sciences2076-34172024-12-0114231137610.3390/app142311376Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro CultureShanshan Zheng0Masato Nakagawa1Yanan Gong2Yasuhiko Matsushima3Satoshi Sasayama4Shunsuke Baba5Yoshitomo Honda6Department of Oral Anatomy, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Oral Anatomy, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Operative Dentistry, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Oral Anatomy, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Oral Implantology, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Oral Implantology, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanDepartment of Oral Anatomy, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata 573-1121, JapanRat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in DPSCs after 3–26 passages (3P–26P). The results show that cell morphology and nuclear size increase proportionally with passage number. The phosphorylated histone H2A.X (γ-H2A.X) positive cells (indicating DNA damage) increased significantly earlier than the 4-Hydroxynonenal (4-HNE) stained cells (indicating an abundance of intracellular reactive oxygen species). Compared to the cells subjected to 3P and 5P, the cells subjected to 15P showed reduced proliferation despite being positive for Ki67. Furthermore, cell growth was completely arrested after 26P. The senescence markers, senescence-associated β-galactosidase (SA-β-gal) and p16, exhibited similar expression patterns that were not correlated with those of p21 and urokinase-type plasminogen activator receptor (uPAR). Nearly all cells expressed SA-β-gal and p16 after 26P, whereas only half expressed p21 and uPAR. These results will contribute to understanding the characteristics of DPSCs toward replicative senescence, which are applicable to elucidate mechanisms related to regenerative medicine and stem cell aging.https://www.mdpi.com/2076-3417/14/23/11376dental pulp stem cellcellular senescencesenescence markerslong-term culture |
| spellingShingle | Shanshan Zheng Masato Nakagawa Yanan Gong Yasuhiko Matsushima Satoshi Sasayama Shunsuke Baba Yoshitomo Honda Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture Applied Sciences dental pulp stem cell cellular senescence senescence markers long-term culture |
| title | Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture |
| title_full | Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture |
| title_fullStr | Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture |
| title_full_unstemmed | Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture |
| title_short | Temporal Changes Toward Cellular Senescence in Rat Dental Pulp Stem Cells Induced by Long-Term In Vitro Culture |
| title_sort | temporal changes toward cellular senescence in rat dental pulp stem cells induced by long term in vitro culture |
| topic | dental pulp stem cell cellular senescence senescence markers long-term culture |
| url | https://www.mdpi.com/2076-3417/14/23/11376 |
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