ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion
Bone marrow stem cells (BMSCs) are a promising source of seed cells in bone tissue engineering, which needs a great quantity of cells. Cell senescence occurs as they are passaged, which could affect the therapeutic effects of cells. Therefore, this study aims to explore the transcriptomic difference...
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MDPI AG
2023-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/11/9766 |
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| author | Yueming Tian Menglong Hu Xuenan Liu Xu Wang Dazhuang Lu Zheng Li Yunsong Liu Ping Zhang Yongsheng Zhou |
| author_facet | Yueming Tian Menglong Hu Xuenan Liu Xu Wang Dazhuang Lu Zheng Li Yunsong Liu Ping Zhang Yongsheng Zhou |
| author_sort | Yueming Tian |
| collection | DOAJ |
| description | Bone marrow stem cells (BMSCs) are a promising source of seed cells in bone tissue engineering, which needs a great quantity of cells. Cell senescence occurs as they are passaged, which could affect the therapeutic effects of cells. Therefore, this study aims to explore the transcriptomic differences among the uncultured and passaged cells, finding a practical target gene for anti-aging. We sorted PαS (PDGFR-α<sup>+</sup>SCA-1<sup>+</sup>CD45<sup>-</sup>TER119<sup>-</sup>) cells as BMSCs by flow cytometry analysis. The changes in cellular senescence phenotype (Counting Kit-8 (CCK-8) assay, reactive oxygen species (ROS) test, senescence-associated β-galactosidase (SA-β-Gal) activity staining, expression of aging-related genes, telomere-related changes and in vivo differentiation potential) and associated transcriptional alterations during three important cell culture processes (in vivo, first adherence in vitro, first passage, and serial passage in vitro) were studied. Overexpression plasmids of potential target genes were made and examed. Gelatin methacryloyl (GelMA) was applied to explore the anti-aging effects combined with the target gene. Aging-related genes and ROS levels increased, telomerase activity and average telomere length decreased, and SA-β-Gal activities increased as cells were passaged. RNA-seq offered that imprinted zinc-finger gene 1 (<i>Zim1</i>) played a critical role in anti-aging during cell culture. Further, <i>Zim1</i> combined with GelMA reduced the expression of P16/P53 and ROS levels with doubled telomerase activities. Few SA-β-Gal positive cells were found in the above state. These effects are achieved at least by the activation of Wnt/β-catenin signaling through the regulation of <i>Wnt2</i>. The combined application of <i>Zim1</i> and hydrogel could inhibit the senescence of BMSCs during in vitro expansion, which may benefit clinical application. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T03:04:16Z |
| publishDate | 2023-06-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-01b01efa879d46d0b4112fd7dc23512b2023-11-18T08:03:44ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-06-012411976610.3390/ijms24119766ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro ExpansionYueming Tian0Menglong Hu1Xuenan Liu2Xu Wang3Dazhuang Lu4Zheng Li5Yunsong Liu6Ping Zhang7Yongsheng Zhou8Department of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaDepartment of Prosthodontics, Peking University School and Hospital of Stomatology, 22 Zhongguancun South Avenue, Haidian District, Beijing 100081, ChinaBone marrow stem cells (BMSCs) are a promising source of seed cells in bone tissue engineering, which needs a great quantity of cells. Cell senescence occurs as they are passaged, which could affect the therapeutic effects of cells. Therefore, this study aims to explore the transcriptomic differences among the uncultured and passaged cells, finding a practical target gene for anti-aging. We sorted PαS (PDGFR-α<sup>+</sup>SCA-1<sup>+</sup>CD45<sup>-</sup>TER119<sup>-</sup>) cells as BMSCs by flow cytometry analysis. The changes in cellular senescence phenotype (Counting Kit-8 (CCK-8) assay, reactive oxygen species (ROS) test, senescence-associated β-galactosidase (SA-β-Gal) activity staining, expression of aging-related genes, telomere-related changes and in vivo differentiation potential) and associated transcriptional alterations during three important cell culture processes (in vivo, first adherence in vitro, first passage, and serial passage in vitro) were studied. Overexpression plasmids of potential target genes were made and examed. Gelatin methacryloyl (GelMA) was applied to explore the anti-aging effects combined with the target gene. Aging-related genes and ROS levels increased, telomerase activity and average telomere length decreased, and SA-β-Gal activities increased as cells were passaged. RNA-seq offered that imprinted zinc-finger gene 1 (<i>Zim1</i>) played a critical role in anti-aging during cell culture. Further, <i>Zim1</i> combined with GelMA reduced the expression of P16/P53 and ROS levels with doubled telomerase activities. Few SA-β-Gal positive cells were found in the above state. These effects are achieved at least by the activation of Wnt/β-catenin signaling through the regulation of <i>Wnt2</i>. The combined application of <i>Zim1</i> and hydrogel could inhibit the senescence of BMSCs during in vitro expansion, which may benefit clinical application.https://www.mdpi.com/1422-0067/24/11/9766stem cellscellular senescencetranscriptomicsWnt pathwayhydrogel |
| spellingShingle | Yueming Tian Menglong Hu Xuenan Liu Xu Wang Dazhuang Lu Zheng Li Yunsong Liu Ping Zhang Yongsheng Zhou ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion International Journal of Molecular Sciences stem cells cellular senescence transcriptomics Wnt pathway hydrogel |
| title | ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion |
| title_full | ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion |
| title_fullStr | ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion |
| title_full_unstemmed | ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion |
| title_short | ZIM1 Combined with Hydrogel Inhibits Senescence of Primary PαS Cells during In Vitro Expansion |
| title_sort | zim1 combined with hydrogel inhibits senescence of primary pαs cells during in vitro expansion |
| topic | stem cells cellular senescence transcriptomics Wnt pathway hydrogel |
| url | https://www.mdpi.com/1422-0067/24/11/9766 |
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