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牙髓间充质干细胞中的DNA损伤：一项研究。

DNA damage in dental pulp mesenchymal stem cells: An study.

作者信息

Aramburú Junior Jaime Sardá, Eilers Treichel Tiago Luis, Lemos Pinto Filho Saulo Tadeu, Gehrke Sergio Alexandre, Machado Alencar Kolinski, Cadoná Francine Carla, Mânica da Cruz Ivana Beatrice, Pippi Ney Luis

机构信息

Graduate Program in Veterinary Medicine, Federal University of Santa Maria, Santa Maria, Brazil.

Biotecnos Research Center, Santa Maria, Rio Grande do Sul, Brazil; Catholic University of Uruguay, Montevideo, Uruguay.

出版信息

Vet Res Forum. 2018 Fall;9(4):293-299. doi: 10.30466/vrf.2018.33083. Epub 2018 Dec 15.

DOI:10.30466/vrf.2018.33083
PMID:30713606
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6346493/
Abstract

The aim of this study was to evaluate the potential use of a DNA comet assay, DNA fragmentation fluorimetric assay and reactive oxygen species levels as potential biomarkers of genome conditions of dental pulp stem cells (DPSCs) isolated from dog canine teeth. Mesenchymal stem cells were isolated from the dental pulp collected from dog teeth. The results obtained suggest the ideal moment for clinical application of cellular therapy for this type of cell. The cell culture was maintained with Dulbecco's modified Eagle's medium supplemented with 10.00% fetal bovine serum for eight passages. During each passage, cell proliferation, oxidative stress and level of DNA fragmentation were assessed by3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium (MTT) assay, testing 2,7 dichlorodihydro-fluorescein-diacetate and PicoGreen, respectively. There were important differences among the first three DPSC passages compared to passages 4-8 and a large number of nuclei with some levels of DNA damage (30.00 to 40.00% in initial DPSC passages and > 50.00% in late passages), indicating DPSC genomic fragility. Within the limitations of this study, the results suggest these relatively simple and inexpensive approaches - comet and DNA fragmentation assays - could help sort stem cells with less DNA damage for use in research or therapies.

摘要

本研究的目的是评估DNA彗星试验、DNA片段化荧光测定法和活性氧水平作为从犬齿分离的牙髓干细胞（DPSC）基因组状况潜在生物标志物的潜在用途。从犬齿收集的牙髓中分离间充质干细胞。所得结果表明了这种类型细胞进行细胞治疗临床应用的理想时机。细胞培养物用补充有10.00%胎牛血清的杜尔贝科改良 Eagle 培养基维持八代。在每一代中，分别通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）试验、2,7-二氯二氢荧光素二乙酸酯和PicoGreen检测来评估细胞增殖、氧化应激和DNA片段化水平。与第4 - 8代相比，前三代DPSC之间存在重要差异，并且有大量细胞核存在一定程度的DNA损伤（初始DPSC代中为30.00%至40.00%，后期代中>50.00%），表明DPSC基因组脆弱性。在本研究的局限性内，结果表明这些相对简单且廉价的方法——彗星试验和DNA片段化测定法——有助于筛选出DNA损伤较小的干细胞用于研究或治疗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/c77b913f05f1/vrf-9-293-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/27e72e76dac5/vrf-9-293-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/2f114d9a8d9a/vrf-9-293-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/263219b9ea9b/vrf-9-293-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/7cd558fc7530/vrf-9-293-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/c77b913f05f1/vrf-9-293-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/27e72e76dac5/vrf-9-293-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/2f114d9a8d9a/vrf-9-293-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/263219b9ea9b/vrf-9-293-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/7cd558fc7530/vrf-9-293-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa9b/6346493/c77b913f05f1/vrf-9-293-g005.jpg

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本文引用的文献

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在体外扩增过程中，肾祖细胞比间充质干细胞具有更高的遗传稳定性和更低的氧化应激。
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