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

doi:10.19439/j.sjos.2018.04.006

上海口腔医学 ›› 2018, Vol. 27 ›› Issue (4): 365-369.doi: 10.19439/j.sjos.2018.04.006

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

林田¹, 赵文青², 陆彦玲¹, 刘玉莹³, 包丽荣³, 吴煜¹

1.广西医科大学附属口腔医院,广西南宁 530021;
2.济宁医学院,山东济宁 272000;
3.广西医科大学研究生院,广西南宁 530021

收稿日期:2017-12-15 修回日期:2018-01-30 出版日期:2018-08-25 发布日期:2018-10-09
通讯作者: 吴煜,E-mail:wuyu0912@163.com
作者简介:林田（1981-）,男,硕士,主治医师, E-mail:8347673@qq.com

Human umbilical cord mesenchymal stem cells co-cultured with dental pulp cells in vitro and its effect on cell biological behaviors

LIN Tian¹, ZHAO Wen-qing², LU Yan-ling¹, LIU Yu-ying³, BAO Li-rong³, WU Yu¹

1.Affiliated Stomatological Hospital of School of Guangxi Medical University. Nanning 530021, Guangxi Zhuang Autonomous Region;
2.Jining Medical University. Jining 272000, Shandong Province;
3.Guangxi Medical University. Naning 530021, Guangxi Zhuang Autonomous Region, China

Received:2017-12-15 Revised:2018-01-30 Online:2018-08-25 Published:2018-10-09

摘要/Abstract

摘要： 目的: 研究人脐带间充质干细胞（human umbilical cord mesenchymal stem cells, hUCMSCs）与经骨形态发生蛋白2（bone morphogenetic protein 2,BMP2）诱导后的人牙髓细胞（human dental pulp cells,hDPCs）共培养对细胞生物学特性的影响。方法: 分别取原代培养的hUCMSCs和hDPCs,分别通过流式细胞术、成骨诱导、成脂诱导以及免疫组织化学染色鉴定细胞;使用BMP2诱导hDPCs,14 d后检查其DSPP、ALP、DMP1的mRNA表达情况;按照1∶1、1∶5、5∶1的比例直接共培养hUCMSCs与经BMP2诱导后的hDPCs,实时定量PCR检测其DSPP、ALP、DMP1、OCN、VEGF、HGF、Nanog的mRNA表达情况。根据实时定量PCR结果选取1∶1组与单独培养hUCMSCs组、hDPCs组比较,分别培养21 d后进行茜素红染色,在酶标仪上于562 nm处检测沉淀物形成情况。采用SPSS21.0软件包对数据进行统计学分析。结果: 直接共培养14 d后,1∶1细胞组的DSPP、ALP、DMP1、OCN、VEGF、HGF的mRNA表达量比单独培养的hUCMSCs组显著升高（P<0.05）,而Nanog mRNA表达量比单独培养的hUCMSCs组降低（P<0.05）。茜素红染色结果显示,1∶1组的OD值显著高于hUCMSCs组（P<0.05）。结论: 将hUCMSCs与经BMP2诱导后的hDPCs按照1∶1比例共培养,可以诱导细胞向成牙本质细胞方向分化,并促进血管生成因子的表达。

关键词: 人脐带间充质干细胞, 牙髓细胞, 直接共培养

Abstract: PURPOSE: To investigated the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) and human dental pulp cells (hDPCs) on cell biological behaviors by co-culture system in vitro. METHODS: hUCMSCs and hDPCs were obtained by primary culture. A culture system of hUCMSCs and hDPCs induced by BMP2 was established in vitro. hUCMSCs and hDPCs were co-cultured at the ratio of 1:1, 1:5 and 5:1. The optimum ratio of each group was selected to further experiment. The formation of calcium nodule was stained by alizarin red staining at 21 day. The expression of DSPP,ALP,DMP1,OCN,VEGF,HGF and Nanog gene was detected by real-time quantitative PCR at 7 day and 14 day. 1:1 group and hUCMSCs, hDPCs group were selected for alizarin red staining at 21 day according to PCR results. Statistical analysis was performed using SPSS 21.0 software package. RESULTS: Calcified nodules formation in 1:1 group was significantly higher than in hUCMSCs group (P<0.05), close to that in hDPCs. qPCR showed that the mRNA expression of DSPP, ALP, DMP1, OCN, VEGF and HGF in 1:1 group was significantly higher than that in hUCMSCs (P<0.05); mRNA expression of Nanog in 1:1 group was significantly lower than in hUCMSCs group (P<0.05). The results of alizarin red staining showed that the OD value of 1:1 group was significantly higher than that of hUCMSCs group (P<0.05). CONCLUSIONS: The cells can be induced to differentiate into odontoblastoid-like cells and the mRNA expression of angiogenic factors was stimulated by hUCMSCs co-culure wih hDPCs.

Key words: Human umbilical cord mesenchymal stem cells, Human dental pulp cells, Co-culture

中图分类号:

R781.3

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.

LIN Tian, ZHAO Wen-qing, LU Yan-ling, LIU Yu-ying, BAO Li-rong, WU Yu. Human umbilical cord mesenchymal stem cells co-cultured with dental pulp cells in vitro and its effect on cell biological behaviors[J]. Shanghai Journal of Stomatology, 2018, 27(4): 365-369.

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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

Human umbilical cord mesenchymal stem cells co-cultured with dental pulp cells in vitro and its effect on cell biological behaviors

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