Pilbauerova Nela, Soukup Tomas, Suchankova Kleplova Tereza, Schmidt Jan, Suchanek Jakub
Department of Dentistry, Charles University, Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, 500 05 Hradec Kralove, Czech Republic.
Department of Histology and Embryology, Charles University, Faculty of Medicine in Hradec Kralove, 500 03 Hradec Kralove, Czech Republic.
Biomolecules. 2021 Mar 20;11(3):464. doi: 10.3390/biom11030464.
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.
端粒是随着每次细胞分裂而缩短的重复性核蛋白DNA序列。干细胞激活端粒酶以补偿端粒损失。本研究旨在评估培养传代对牙髓干细胞相对端粒长度和增殖能力的影响。我们使用了10个通过非控制速率冷冻保存12个月的牙髓干细胞（DPSC）谱系来实现该研究目标。我们分析了它们的增殖率、使用流式细胞术检测表型、多能性以及使用qPCR分析检测相对端粒长度。在未传代的培养1周、2周和3周后进行分析，从而在补充研究中确定相对端粒长度。我们记录了随着传代增加的端粒损耗。在培养结束时观察到，相对端粒长度越短，达到的群体倍增数越低，群体倍增时间越长。在随后的补充研究中，我们观察到未传代培养两周的DPSC中端粒延长。我们得出结论，体外培养过程中对DPSC的过度增殖需求会导致端粒损耗。我们提出了这样一种理论，即在未传代的细胞培养过程中端粒酶可能更有效。这一观察结果有助于在体外DPSC扩增过程中保持端粒长度。
