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Expression of senescence markers in human PDL stem cells after long-term cultivation in vitro
Biotechnology & Biotechnological Equipment ( IF 1.4 ) Pub Date : 2021-07-09 , DOI: 10.1080/13102818.2021.1946427
Zornitsa Mihaylova 1 , Marina Miteva 2 , Pavel Stanimirov 1 , Evgeniy Aleksiev 1 , Vanyo Mitev 2 , Nikolay Ishkitiev 2
Affiliation  

Abstract

Cell senescence is a continuous irreversible process, ending with cell death. Long-term in vitro cultivation leads to decreased proliferation ability, disruption in cellular morphology and function in somatic cells. The aim of the present study was to investigate the markers of cell cycle arrest and senescence after long-term cultivation of periodontal ligament stem cells (PDLSC). The cells were isolated from routinely extracted third molars via enzymatic digestion and cultured continuously up to passage 16. Cell count and population doubling were evaluated at each passage. The enzymatic activity of telomerase and β-galactosidase were assessed, as these are well-known markers for cell senescence and aging. The results demonstrated two peaks with significantly increased cellular proliferation rate at passage 6 and passage 12. Slight differences in the proliferative ability were identified between cells from 1st and 16th passages without any statistical significance. Significant decrease in telomerase activity was observed starting immediately after the first passaging. β-Galactosidase activity was found to be uninterrupted following long-term in vitro cultivation. Our study indicates that the PDL stem cells did not show a significant decrease in the proliferation ability up to the 16th passage.



中文翻译:

体外长期培养人PDL干细胞衰老标志物的表达

摘要

细胞衰老是一个持续的不可逆过程,以细胞死亡告终。长期体外培养导致增殖能力下降、细胞形态和体细胞功能受到破坏。本研究的目的是研究牙周膜干细胞 (PDLSC) 长期培养后细胞周期停滞和衰老的标志物。将细胞从常规提取第三磨牙中分离通过酶消化并连续培养至第 16 代。在每次传代时评估细胞计数和群体倍增。评估了端粒酶和 β-半乳糖苷酶的酶活性,因为它们是众所周知的细胞衰老和衰老标志物。结果表明,在第 6 代和第 12 代细胞增殖率显着增加的两个峰。在第 1 代和第 16 代细胞之间鉴定出增殖能力略有差异,没有任何统计学意义。在第一次传代后立即观察到端粒酶活性显着降低。长期体外试验后发现β-半乳糖苷酶活性不受干扰栽培。我们的研究表明,直到第 16 代,PDL 干细胞的增殖能力并没有显着下降。

更新日期:2021-07-09
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