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  

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

作　　者：杨峥[1] 赵敏 肖琳 陈思萍 YANG Zheng;ZHAO Min;XIAO Lin;CHEN Si-ping(Department of Stomatology,Jiangmen Maternal and Child Healh Care Haspial Jiangmen 5900;Deartment of Stomaologs,Stomatological Hospial of Southem Medical University Guangzhou 510000,Guangdong Province,China)

机构地区：[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

出　　处：《上海口腔医学》2021年第3期247-252,共6页Shanghai Journal of Stomatology

基　　金：广东省医学科研资助项目(A2017618)。

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%O_(2)(3%低氧浓度实验组)、5%O_(2)(5%低氧浓度实验组)和21%O_(2)(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%O_(2)条件下培养4.7d,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。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 mea-sured 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 were97.25%, 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.

关 键 词：低氧浓度 体外培养 生物学特性 人牙髓干细胞 

分 类 号：Q256[生物学—细胞生物学]