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Passage Effect on Aging of Human Umbilical Cord Derived Mesenchymal Stem Cell

机译:人脐带间充质干细胞衰老的传代效应

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摘要

To be used in regenerative medicine, cells should be checked for various conditions, including cell aging. This study aimed to learn senescent profile and its relation to cell viability, proliferation and cell size in various passages, which were done in α-MEM-10% PRP medium, until senescence-associated β-galactosidase (SA-β-Gal) positive cells were found. Stem cells were isolated from umbilical cord tissue by multiple harvest explant method, cultured in α-MEM-10% PRP until P-17 and stained using SA-β-Gal staining. Viability, Population Doubling Time (PDT), percentage SA-β-Gal (+) and cell size at 30% confluent and at senescent staining were analyzed. Passages with SA-β-Gal (+) and (-) were compared in term of viability, PDT and cell size at 30% confluent. Further, cell size at senescent staining between SA-β-Gal (+) and (-) groups were compared. Viability and PDT showed no significant difference between SA-β-Gal (+) and (-) groups, while cell size at 30% confluence showed significant increase in SA-β-Gal (+) compared to (-) groups. Further, cell size in senescent staining showed significantly smaller cell size in SA-β-Gal (-) compared to SA-β-Gal (+) cells. Moreover, this study showed that even in SA-β-Gal (+) group, viability was greater than 91%, PDT was less than 2.1 days and cell size was less than 2602 µm2. In conclusion, umbilical cord derived MSCs that were cultured in α-MEM-10% PRP began to undergo aging at P-10. Morphological criteria of UC-MSC aging were cell size greater than 2602 µm2 with a change in morphology toward a rounded shape.
机译:要用于再生医学,应检查细胞的各种状况,包括细胞老化。这项研究旨在了解衰老过程及其在不同传代中与细胞活力,增殖和细胞大小的关系,这些过程在α-MEM-10%PRP培养基中进行,直到衰老相关的β-半乳糖苷酶(SA-β-Gal)阳性找到了细胞。通过多次收获外植体方法从脐带组织中分离出干细胞,在α-MEM-10%PRP中培养直至P-17,然后使用SA-β-Gal染色。分析了30%融合和衰老染色时的活力,种群倍增时间(PDT),SA-β-Gal(+)百分比和细胞大小。比较了SA-β-Gal(+)和(-)在30%汇合时的存活率,PDT和细胞大小方面的传代。此外,比较了SA-β-Gal(+)和(-)组之间在衰老染色时的细胞大小。与(-)组相比,SA-β-Gal(+)和(-)组之间的活力和PDT没有显着差异,而在30%汇合时,细胞大小显示SA-β-Gal(+)组的显着增加。此外,与SA-β-Gal(+)细胞相比,衰老染色中的细胞大小显示SA-β-Gal(-)细胞显着更小。此外,该研究表明,即使在SA-β-Gal(+)组中,生存力也大于91%,PDT小于2.1天,细胞大小小于2602 µm2。总之,在α-MEM-10%PRP中培养的来自脐带的MSC在P-10处开始老化。 UC-MSC老化的形态学标准是细胞大小大于2602 µm2,形态向圆形变化。

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