上海口腔医学 ›› 2021, Vol. 30 ›› Issue (3): 247-252.doi: 10.19439/j.sjos.2021.03.005

• 论著 • 上一篇    下一篇

不同低氧浓度对体外培养人牙髓干细胞生物学特性的影响

杨峥1, 赵敏1, 肖琳1, 陈思萍2   

  1. 1.江门市妇幼保健院 口腔科,广东 江门 529000;
    2.南方医科大学口腔医院 口腔科,广东 广州 510000
  • 收稿日期:2020-05-15 修回日期:2020-08-19 出版日期:2021-06-25 发布日期:2021-08-05
  • 通讯作者: 陈思萍,E-mail:1145265689@qq.com
  • 作者简介:杨峥(1983-),女,本科,主治医师,E-mail:yc198417@163.com
  • 基金资助:
    广东省医学科研资助项目(A2017618)

Effects of different hypoxic concentrations on biological characteristics of human dental pulp stem cells in vitro

YANG Zheng1, ZHAO Min1, XIAO Lin1, CHEN Si-ping2   

  1. 1. Department of Stomatology, Jiangmen Maternal and Child Health Care Hospital. Jiangmen 529000;
    2. Department of Stomatology, Stomatological Hospital of Southern Medical University. Guangzhou 510000, Guangdong Province, China
  • Received:2020-05-15 Revised:2020-08-19 Online:2021-06-25 Published:2021-08-05

摘要: 目的: 探讨不同低氧浓度对体外培养人牙髓干细胞生物学特性的影响。方法: 选择2019年1月—2020年1月收集的健康者完整拔除的下颌阻生第三磨牙,采用块酶消化法培养牙髓干细胞,置于3% O2(3%低氧浓度实验组)、5% O2(5%低氧浓度实验组)和21% O2(21%常氧浓度对照组)中培养7 d。用流式细胞仪检测细胞周期、凋亡情况、细胞表面标志物,采用CCK-8法检测细胞增殖情况, Transwell实验检测细胞迁移能力。采用SPSS 20.0软件包对数据进行统计学分析。结果: 传代培养细胞干细胞表面标志物CD44、CD29、D73表达率分别为97.25%、99.36%和99.60%;分别置于3%、5%和21% O2条件下培养4、7 d,5%低氧浓度实验组牙髓干细胞增殖最强,3%低氧浓度实验组牙髓干细胞增殖最弱,差异显著(P<0.05),牙髓干细胞凋亡情况相比无显著差异(P>0.05);21%常氧浓度对照组牙髓干细胞G1期占比显著低于3%和5%低氧浓度实验组(P<0.05),S期细胞占比显著高于3%和5%低氧浓度实验组(P<0.05);3%低氧浓度实验组牙髓干细胞迁移细胞最多,21%常氧浓度对照组牙髓干细胞迁移细胞最少,差异显著(P<0.05)。结论: 不同氧浓度下培养人牙髓干细胞,对其形态、增殖能力、迁移能力、细胞周期影响较大,但对细胞凋亡影响不大。

关键词: 低氧浓度, 体外培养, 生物学特性, 人牙髓干细胞

Abstract: PURPOSE: To investigate the effects of different hypoxic concentrations on biological characteristics of human dental pulp stem cells in vitro. METHODS: Impacted mandibular third molars were extracted from healthy individuals, and the dental pulp stem cells were cultured by tissue block enzyme digestion. Cells cultured under the conditions of 3%, 5% and 21% oxygen concentration for 7 days were set as 3% hypoxia group, 5% hypoxia group, and 21% nomoxia group, respectively. Flow cytometry was used to detect cell surface markers, cell cycle and apoptosis. Cell proliferation was measured by CCK-8 method. Transwell chamber assay was used to detect migration ability. Statistical analysis was completed by SPSS 20.0 software package. RESULTS: The expression rates of CD44, CD29 and D73 of the subculture cells were 97.25%, 99.36% and 99.60%, respectively. The proliferation ability of dental pulp stem cells was the strongest in 5% hypoxia group, and weakest in 3% hypoxia group, with significant difference(P<0.05). The apoptosis rate had no significant difference among various concentrations of oxygen(P>0.05). Compared with 21% nomoxia group, the proportion of dental pulp stem cells in G1 phase was significantly lower than that in 3% hypoxia group and 5% hypoxia group(P<0.05), and cell in S phase was significantly higher than that in 3% hypoxia group and 5% hypoxia group(P<0.05). The migration ability was the strongest in 3% hypoxia group, and weakest in 21% nomoxia group, with significant difference(P<0.05). CONCLUSIONS: Different concentrations of hypoxia have great influence on the morphology, proliferation, migration and cell cycle of human dental pulp stem cells in vitro with little impact on cell apoptosis.

Key words: Hypoxic concentration, In vitro culture, Human dental pulp stem cells, Biological characteristics

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