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.

参考文献

[1] Jiang L, Peng WW, Li LF, et al.Proliferation and multilineage potential of CXCR4-positive human dental pulp cells in vitro [J]. J Endod, 2012, 38(5): 642-647.
[2] Tu MG, Ho CC, Hsu TT, et al.Mineral trioxide aggregate with mussel-inspired surface nanolayers for stimulating odontogenic differentiation of dental pulp cells[J]. J Endod, 2018, 44(6): 963-970.
[3] Yokose S, Naka T.Lymphocyte enhancer-binding factor 1: an essential factor in odontoblastic differentiation of dental pulp cells enzymatically isolated from rat incisors[J]. J Bone Miner Metab, 2010, 28(6): 650-658.
[4] Shimabukuro Y, Ueda M, Ozasa M, et al.Fibroblast growth factor-2 regulates the cell function of human dental pulp cells[J]. J Endod, 2009, 35(11): 1529-1535.
[5] Trubiani O, Caputi S, Di ID, et al.The cytotoxic effects of resin-based sealers on dental pulp stem cells[J]. Int Endod J, 2010, 43(8): 646-653.
[6] Paranjpe A, Zhang H, Johnson JD.Effects of mineral trioxide aggregate on human dental pulp cells after pulp-capping procedures[J]. J Endod, 2010, 36(6): 1042-1047.
[7] Li S, Lin C, Zhang J, et al.Quaking promotes the odontoblastic differentiation of human dental pulp stem cells[J]. J Cell Physiol, 2018, 233(9): 7292-7304.
[8] Paim A, Braghirolli DI, NSM C, et al.Human dental pulp stem cell adhesion and detachment in polycaprolactone electrospun scaffolds under direct perfusion[J]. Braz J Med Biol Res, 2018, 51(5): e6754.
[9] 覃乙芯, 吴卓敏, 徐茜, 等. 血清饥饿对原代人脐静脉内皮细胞周期同步化的影响[J]. 南方医科大学学报, 2016, 36(8): 1140-1143.
[10] Khammanit R, Chantakru S, Kitiyanant Y, et al.Effect of serum starvation and chemical inhibitors on cell cycle synchronization of canine dermal fibroblasts[J]. Theriogenology, 2008, 70(1): 27-34.
[11] 刘斐, 吴补领, 高杰, 等. 血清饥饿法对人牙髓细胞周期同步化的研究[J]. 牙体牙髓牙周病学杂志, 2011, 21(2): 67-71.
[12] 江龙, 朱亚琴. 改良组织块法体外培养人牙周膜细胞和牙髓细胞[J]. 牙体牙髓牙周病学杂志, 2008, 18(4): 195-199.
[13] Peng W, Liu W, Zhai W, et al.Effect of tricalcium silicate on the proliferation and odontogenic differentiation of human dental pulp cells[J]. J Endod, 2011, 37(9): 1240-1246.
[14] 宋春娇, 吕冰洁, 张小玲, 等. 血清饥饿法用于细胞周期同步化的方法学研究[J]. 中国地方病学杂志, 2003, 22(4): 362-364.
[15] 董关木. 细胞培养用牛血清现状与进展[J]. 中国生物制品学杂志, 2007, 20(9): 697-700.
[16] 严丽萍, 陶茂萱. 血清饥饿和接触抑制两种GO期同步化方法效果评价[J]. 卫生研究, 2007, 36(3): 275-278.
[17] De Barros FR, Goissis MD, Caetano HV, et al.Serum starvation and full confluency for cell cycle synchronization of domestic cat (felis catus) foetal fibroblasts[J]. Reprod Domest Anim, 2010, 45(1): 38-41.
[18] 邵晓云, 徐绍业. 血清饥饿法处理胎儿成纤维细胞周期同步化的研究[J]. 中国现代医学杂志, 2010, 20(1): 18-21.
[19] 彭伟伟, 翟万银, 江龙, 等. 人牙髓细胞复合不同孔径羟基磷灰石/磷酸三钙支架材料的生物学行为[J]. 中国组织工程研究与临床康复, 2010, 14(51): 9567-9571.

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 12

编辑推荐

Metrics

本文评价

[1]	张安妮, 丁灿灿, 黄丽苹, 李适廷. 抑制连接蛋白43介导半通道活性促进脂多糖诱导的人牙髓细胞成牙本质分化[J]. 上海口腔医学, 2024, 33(1): 22-29.
[2]	郭梦瑶, 李凯玉, 聂敏海, 刘旭倩. 上调Decorin对口腔鳞癌荷瘤裸鼠EGFR、C-Myc、p21表达的影响[J]. 上海口腔医学, 2023, 32(1): 40-46.
[3]	徐方芳, 蒋备战. DKK1对脂多糖感染人牙髓细胞生物学行为的影响[J]. 上海口腔医学, 2021, 30(4): 355-359.
[4]	曹颖, 彭伟伟, 王璇, 伍甜甜, 杜嵘, 朱亚琴. 重组人结缔组织生长因子对牙髓细胞增殖与成牙本质分化的影响[J]. 上海口腔医学, 2020, 29(1): 7-12.
[5]	林田, 赵文青, 陆彦玲, 刘玉莹, 包丽荣, 吴煜. 人脐带间充质干细胞与牙髓细胞体外共培养对细胞生物学的影响[J]. 上海口腔医学, 2018, 27(4): 365-369.
[6]	李莉芬, 文扬, 江龙, 朱亚琴. 衣霉素诱导牙髓细胞内质网应激模型的建立[J]. 上海口腔医学, 2018, 27(2): 135-138.
[7]	包丽荣, 赵文青, 林田, 陆彦玲, 吴煜. BMP-2诱导人牙髓细胞分化过程中miRNA表达谱的变化[J]. 上海口腔医学, 2017, 26(5): 476-483.
[8]	林泓磊, 江磊, 张长源, 林东红, 程辉. 反复熔铸对钴铬烤瓷合金细胞相容性的影响[J]. 上海口腔医学, 2017, 26(3): 246-250.
[9]	何璇,韦维,陈文霞. 富血小板纤维蛋白凝胶三维结构及其对人牙髓细胞体外增殖的影响[J]. 上海口腔医学, 2015, 24(3): 263-268.
[10]	王艳丽, 潘克清, 孙艳, 邓婧. 脂多糖对大鼠牙髓细胞ALP、BSP、DSPP表达的影响[J]. 上海口腔医学, 2014, 23(4): 431-435.
[11]	伍甜甜;杜嵘;李莉芬;朱亚琴. 应用流式细胞术检测人牙髓细胞内活性氧的方法探讨 [J]. 上海口腔医学, 2012, 21(6): 617-620.
[12]	魏福兰;耿杰;王春玲;王惠;张本君;张凡 . 牵张力作用下人牙髓细胞HIF-1α和VEGF的表达 [J]. 上海口腔医学, 2012, 21(5): 501-505.