The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells
Telomeres are repetitive nucleoprotein DNA sequences that shorten with each cell division. The stem cells activate telomerase to compensate for the telomere loss. This study aimed to evaluate the effect of cultivation passaging on the relative telomere length and proliferation capacity of dental pul...
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MDPI AG
2021-03-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/11/3/464 |
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| author | Nela Pilbauerova Tomas Soukup Tereza Suchankova Kleplova Jan Schmidt Jakub Suchanek |
| author_facet | Nela Pilbauerova Tomas Soukup Tereza Suchankova Kleplova Jan Schmidt Jakub Suchanek |
| author_sort | Nela Pilbauerova |
| collection | DOAJ |
| description | Telomeres are repetitive nucleoprotein DNA sequences that shorten with each cell division. The stem cells activate telomerase to compensate for the telomere loss. This study aimed to evaluate the effect of cultivation passaging on the relative telomere length and proliferation capacity of dental pulp stem cells. We used ten dental pulp stem cell (DPSC) lineages stored for 12 months using uncontrolled-rate freezing to reach the study’s goal. We analyzed their proliferation rate, phenotype using flow cytometry, multipotency, and relative telomere length using a qPCR analysis. We determined the relative telomere length in the added study by performing analysis after one, two, and three weeks of cultivation with no passaging. We documented the telomere attrition with increasing passaging. The shorter the relative telomere length, the lower reached population doublings, and longer population doubling time were observed at the end of the cultivation. We observed the telomere prolongation in DPSCs cultivated for two weeks with no passaging in the added subsequent study. We concluded that excessive proliferation demands on DPSCs during in vitro cultivation result in telomere attrition. We opened the theory that the telomerase might be more efficient during cell cultivation with no passaging. This observation could help in preserving the telomere length during ex vivo DPSC expansion. |
| first_indexed | 2024-03-10T13:03:22Z |
| format | Article |
| id | doaj.art-e83441514e1d4cb3a68fb9b7fb263195 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T13:03:22Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-e83441514e1d4cb3a68fb9b7fb2631952023-11-21T11:20:59ZengMDPI AGBiomolecules2218-273X2021-03-0111346410.3390/biom11030464The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem CellsNela Pilbauerova0Tomas Soukup1Tereza Suchankova Kleplova2Jan Schmidt3Jakub Suchanek4Department of Dentistry, Charles University, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, 500 05 Hradec Kralove, Czech RepublicDepartment of Histology and Embryology, Charles University, Faculty of Medicine in Hradec Kralove, 500 03 Hradec Kralove, Czech RepublicDepartment of Dentistry, Charles University, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, 500 05 Hradec Kralove, Czech RepublicDepartment of Dentistry, Charles University, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, 500 05 Hradec Kralove, Czech RepublicDepartment of Dentistry, Charles University, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, 500 05 Hradec Kralove, Czech RepublicTelomeres are repetitive nucleoprotein DNA sequences that shorten with each cell division. The stem cells activate telomerase to compensate for the telomere loss. This study aimed to evaluate the effect of cultivation passaging on the relative telomere length and proliferation capacity of dental pulp stem cells. We used ten dental pulp stem cell (DPSC) lineages stored for 12 months using uncontrolled-rate freezing to reach the study’s goal. We analyzed their proliferation rate, phenotype using flow cytometry, multipotency, and relative telomere length using a qPCR analysis. We determined the relative telomere length in the added study by performing analysis after one, two, and three weeks of cultivation with no passaging. We documented the telomere attrition with increasing passaging. The shorter the relative telomere length, the lower reached population doublings, and longer population doubling time were observed at the end of the cultivation. We observed the telomere prolongation in DPSCs cultivated for two weeks with no passaging in the added subsequent study. We concluded that excessive proliferation demands on DPSCs during in vitro cultivation result in telomere attrition. We opened the theory that the telomerase might be more efficient during cell cultivation with no passaging. This observation could help in preserving the telomere length during ex vivo DPSC expansion.https://www.mdpi.com/2218-273X/11/3/464telomeretelomerasedental pulp stem cellsqPCRrelative telomere length measurement |
| spellingShingle | Nela Pilbauerova Tomas Soukup Tereza Suchankova Kleplova Jan Schmidt Jakub Suchanek The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells Biomolecules telomere telomerase dental pulp stem cells qPCR relative telomere length measurement |
| title | The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells |
| title_full | The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells |
| title_fullStr | The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells |
| title_full_unstemmed | The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells |
| title_short | The Effect of Cultivation Passaging on the Relative Telomere Length and Proliferation Capacity of Dental Pulp Stem Cells |
| title_sort | effect of cultivation passaging on the relative telomere length and proliferation capacity of dental pulp stem cells |
| topic | telomere telomerase dental pulp stem cells qPCR relative telomere length measurement |
| url | https://www.mdpi.com/2218-273X/11/3/464 |
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