中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (21): 3357-3363.doi: 10.3969/j.issn.2095-4344.1765

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

不同来源间充质干细胞条件培养基对内源性衰老细胞作用的比较

郑桂纯，赵姝灿，黄丽贞，张晓芳，王丙云，陈胜锋，陈志胜   

  1. 佛山科学技术学院生命科学与工程学院，广东省佛山市 528200
  • 修回日期:2019-03-24 出版日期:2019-07-28 发布日期:2019-07-28
  • 通讯作者: 陈志胜，教授，博士，硕士生导师，佛山科学技术学院生命科学与工程学院，广东省佛山市 528200
  • 作者简介:郑桂纯，女，1994年生，广东省普宁市人，汉族，在读硕士，主要从事干细胞研究。 并列第一作者：赵姝灿，女，1995年生，广东省佛山市人，汉族，在读硕士，主要从事干细胞研究。
  • 基金资助:

    广东省自然科学基金项目(2018A030313892)，项目负责人：陈胜锋；广东省教育厅预防兽医学重点实验室项目(2014KTSPT037)，项目参与人：王丙云；干细胞基础研究与应用开放平台(佛财工[2015]143号)，项目负责人：陈志胜

Effects of mesenchymal stem cell conditioned media from different sources on intrinsic aging cells

Zheng Guichun, Zhao Shucan, Huang Lizhen, Zhang Xiaofang, Wang Bingyun, Chen Shengfeng, Chen Zhisheng   

  1. College of Life Science and Engineering, Foshan University, Foshan 528200, Guangdong Province, China
  • Revised:2019-03-24 Online:2019-07-28 Published:2019-07-28
  • Contact: Chen Zhisheng, MD, Professor, Master’s supervisor, College of Life Science and Engineering, Foshan University, Foshan 528200, Guangdong Province, China
  • About author:Zheng Guichun, Master candidate, College of Life Science and Engineering, Foshan University, Foshan 528200, Guangdong Province, China; Zhao Shucan, Master candidate, College of Life Science and Engineering, Foshan University, Foshan 528200, Guangdong Province, China. Zheng Guichun and Zhao Shucan contributed equally to this work.
  • Supported by:

    the Natural Science Foundation of Guangdong Province, No. 2018A030313892 (to CSF); Key Laboratory Project of Preventive Veterinary Medicine, Guangdong Provincial Department of Education, No. 2014KTSPT037 (to WBY); Stem Cell Basic Research and Application Open Platform, No. [2015]143 (to CZS)

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摘要：

文章快速阅读：

文题释义：
活性氧：正常情况下细胞内活性氧的产生和清除处于平衡，但在细胞衰老进程或刺激因子存在时，其胞内清除活性氧的调节机制失衡，活性氧可诱发膜脂过氧化、蛋白质过氧化、DNA损伤等，导致细胞结构和功能受损。
KLOTHO基因：被誉为衰老抑制基因，当其表达量上调时可以延缓人体衰老，当其表达量下调时会出现类似于衰老的表现。

 

摘要
背景：随着经济的发展，人们对美的追求越来越高，抗衰老研究成为当今的研究热点。干细胞是具有自我更新能力和多向分化能力的细胞，间充质干细胞是其中的一种。研究表明间充质干细胞能通过旁分泌作用修复损伤衰老的细胞，但对于不同来源间充质干细胞条件培养基的抗皮肤衰老作用研究甚少。
目的：比较人脂肪、羊水、胎盘、脐带来源间充质干细胞条件培养基对自然衰老皮肤成纤维细胞的抗衰老作用。
方法：制备人脂肪、羊水、胎盘、脐带4种不同来源的间充质干细胞条件培养基，添加到自然衰老皮肤成纤维细胞中，采用CCK-8法检测细胞活力、β-半乳糖苷酶染色试剂盒检测细胞衰老率、DCFH-DA探针检测活性氧含量以及qPCR检测p16、p53、COL1、KLOTHO基因表达量。
结果与结论：①4种间充质干细胞条件培养基均能提高皮肤成纤维细胞的活力，其中脂肪间充质干细胞条件培养基的效果最好，β-半乳糖苷酶染色率最低；②4种间充质干细胞条件培养基均能降低活性氧含量，脂肪和羊水来源间充质干细胞条件培养基组的活性氧含量最低；③p16和p53在胎盘和脐带间充质干细胞条件培养基组的表达均高于对照组，在羊水来源间充质干细胞条件培养基组的表达较低，脂肪间充质干细胞条件培养基组p16表达显著低于对照组，但p53的表达较高；④COL1基因表达除羊水间充质干细胞条件培养基组外，其他3组均显著高于对照组，KLOTHO基因表达除胎盘间充质干细胞条件培养基组外，其他3组均显著高于对照组，且脂肪间充质干细胞条件培养基组的表达最高；⑤结果显示，4种不同来源间充质干细胞条件培养基能在一定程度上延缓细胞衰老，但作用效果存在差异，其作用机制还有待进一步研究。


中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-3017-2546(郑桂纯)

关键词: 皮肤成纤维细胞, 内源性衰老, 脂肪间充质干细胞, 胎盘间充质干细胞, 脐带间充质干细胞, 羊水间充质干细胞, 间充质干细胞条件培养基, 广东省自然科学基金

Abstract:

 

BACKGROUND: With the economic development, people have an increasing pursuit of beauty, and anti-aging research has become a hot research topic. Stem cells have self-renewal and multi-directional differentiation ability, and mesenchymal stem cells are one of them. Studies have shown that mesenchymal stem cells can repair aging cells through paracrine action, but little has been reported on the anti-aging effects of mesenchymal stem cell conditioned media from different sources.
OBJECTIVE: To compare the anti-aging effects of human adipose-, amniotic fluid-, placenta- and umbilical cord-derived mesenchymal stem cell conditioned media on aging skin fibroblasts. 
METHODS: Human adipose, amniotic fluid, placenta and umbilical cord-derived mesenchymal stem cell conditioned media were prepared and used to culture aging skin fibroblasts. Cell viability was detected by cell counting kit-8 and cell senescence rate measured by β-galactosidase staining kit. DCFH-DA probe was used to detect the content of reactive oxygen species. qPCR was used to detect the expression levels of p16, p53, COL1 and KLOTHO genes. 
RESULTS AND CONCLUSION: (1) Four sources of mesenchymal stem cell conditioned media could increase the cell viability of skin fibroblasts, among which the adipose-derived mesenchymal stem cell conditioned medium had the best effect and the lowest β-galactosidase staining rate. (2) Four sources of mesenchymal stem cell conditioned media could reduce reactive oxygen species content, and adipose- and amniotic fluid-derived mesenchymal stem cell conditioned media had the lowest content of reactive oxygen species. (3) p16 and p53 levels in the placenta and umbilical cord-derived mesenchymal stem cell conditioned media were higher than those in the control group. The expression of p53 in the amniotic fluid-derived mesenchymal stem cell conditioned medium was lower than that in the control group, but there was no difference in the p16 expression between the two groups. Compared with the control group, the expression of p16 was lower and the expression of p53 was higher in the adipose-derived mesenchymal stem cell conditioned medium. (4) Compared with the control group, the expression of COL1 gene was significantly higher in the different culture media except for the amniotic fluid-derived mesenchymal stem cell conditioned medium. Compared with the control group, the expression of KLOTHO was significantly higher in different culture media except for placenta-derived mesenchymal stem cell conditioned medium, and highest in the adipose-derived mesenchymal stem cell conditioned medium. To conclude, mesenchymal stem cell conditioned media from four different sources can delay cell senescence to a certain extent, though its mechanism remains to be further studied.

Key words: skin fibroblasts, endogenous senescence, adipose-derived mesenchymal stem cells, placenta-derived mesenchymal stem cells, umbilical cord mesenchymal stem cells, amniotic fluid mesenchymal stem cells, mesenchymal stem cell conditioned medium, Natural Science Foundation of Guangdong Province

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