血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响

doi:10.19439/j.sjos.2019.01.001

上海口腔医学 ›› 2019, Vol. 28 ›› Issue (1): 1-5.doi: 10.19439/j.sjos.2019.01.001

血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响

彭伟伟, 戴兆威, 曹颖, 韩俊力^*, 朱亚琴^*

上海交通大学医学院附属第九人民医院·口腔医学院口腔综合科,国家口腔疾病临床医学研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海 200011

收稿日期:2018-09-07 修回日期:2018-11-10 出版日期:2019-02-25 发布日期:2019-04-12
通讯作者: 朱亚琴,E-mail：zyq1590@163.com;韩俊力,E-mail：hanjunli@sina.com。^*共同通信作者
作者简介:彭伟伟（1984-）,女,硕士,住院医师,E-mail:pww1027@163.com
基金资助:
国家自然科学基金（81700949）; 高等学校博士学科点专项科研基金（20130073110013）; 上海高校高峰高原学科建设项目; 上海市口腔医学研究所院级基金（2017-08）

Effect of serum starvation and culture to confluence on cell cycle synchronization and mineralization of human dental pulp cells

PENG Wei-wei, DAI Zhao-wei, CAO Ying, HAN Jun-li, ZHU Ya-qin

Department of General Dentistry, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology and Shanghai Research Institute of Stomatology. Shanghai 200011, China

Received:2018-09-07 Revised:2018-11-10 Online:2019-02-25 Published:2019-04-12

摘要/Abstract

摘要： 目的:比较血清饥饿及融合培养对人牙髓细胞周期同步化及矿化活性的影响。方法:将人牙髓细胞分别培养至80%及100%融合后,使用含0.5%胎牛血清的培养基继续培养细胞24、48、72 h,使用流式细胞仪检测牙髓细胞的细胞周期。以100%融合培养后饥饿48 h的人牙髓细胞作为实验组,80%融合培养的细胞作为对照组,在基因水平检测碱性磷酸酶、Ⅰ型胶原、骨钙素的表达;在蛋白水平检测碱性磷酸酶活性。采用SPSS13.0软件包对数据进行统计学分析。结果:在人牙髓细胞达到100%融合后,经血清饥饿48 h的G0/G1期细胞比100%融合培养组以及血清饥饿组多（P<0.05）。在基因水平,实验组Ⅰ型胶原、骨钙素的表达与对照组无统计学差异, 但是能促进碱性磷酸酶的表达（P<0.05）,同时在蛋白水平也刺激了人牙髓细胞碱性磷酸酶的分泌（P<0.05）。结论:100%融合培养联合血清饥饿法使更多的人牙髓细胞周期同步于G0/G1期,能更好地促进人牙髓细胞矿化。

关键词: 牙髓细胞, 血清饥饿, 融合培养, 细胞周期

Abstract: PURPOSE: To compare the effect of serum starvation and culture to confluence on cell cycle synchronization and mineralization of human dental pulp cells (hDPCs). METHODS: HDPCs were cultured to 80% and 100% confluence respectively, and then cultured for 24, 48 and 72 hours by culture medium containing 0.5% fetal bovine serum(FBS). Cell cycle of hDPCs were identified by flow cytometry. Then hDPCs cultured by serum starvation for 48h after culturing to 100% confluence were used as the experimental group, and hDPCs cultured to 80% confluence were used as the control group. The expression of alkaline phosphatase(ALP), collagen type Ⅰ(COL-Ⅰ) and osteocalcin(OCN) was detected at gene level; activity of ALPase was detected at protein level. SPSS 13.0 software was used for statistical analysis. RESULTS: When hDPCs were cultured by serum starvation for 48h after culturing to 100% confluence, cells at G0/G1 stage were more than culture to 100% confluence and serum starvation group (P<0.05). At the genetic level, the expression of COL-Ⅰand OC in the experimental group was not statistically different from that of the control group, but can promote the expression of ALP(P<0.05), and stimulate the secretion of hDPCs at protein level at the same time (P<0.05). CONCLUSIONS: Culture to confluence combined serum starvation can synchronize more hDPCs at G0/G1 stage and promote mineralization of hDPCs.

Key words: Dental pup cell, Serum starvation, Culture to confluence, Cell cycle

中图分类号:

R781.3

彭伟伟, 戴兆威, 曹颖, 韩俊力, 朱亚琴. 血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响[J]. 上海口腔医学, 2019, 28(1): 1-5.

PENG Wei-wei, DAI Zhao-wei, CAO Ying, HAN Jun-li, ZHU Ya-qin. Effect of serum starvation and culture to confluence on cell cycle synchronization and mineralization of human dental pulp cells[J]. Shanghai Journal of Stomatology, 2019, 28(1): 1-5.

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血清饥饿及融合培养对人牙髓细胞周期同步化和矿化活性的影响

Effect of serum starvation and culture to confluence on cell cycle synchronization and mineralization of human dental pulp cells

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