[1]余勇,陶昱,邓锋,等.体外共培养人牙髓干细胞对人牙囊干细胞增殖、成骨分化的作用[J].陆军军医大学学报(原第三军医大学学报),2015,37(22):2267-2272.
 Yu Yong,Tao Yu,Deng Feng,et al.Role of in vitro co-culture of human dental pulp stem cells in proliferation and osteogenic differentiation of human dental follicle stem cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(22):2267-2272.
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第22期
页码:
2267-2272
栏目:
论著
出版日期:
2015-11-30

文章信息/Info

Title:
Role of in vitro co-culture of human dental pulp stem cells in proliferation and osteogenic differentiation of human dental follicle stem cells
作者:
余勇陶昱邓锋王睿陈军
重庆医科大学附属口腔医院正畸科,口腔疾病与生物医学重庆市重点实验室,重庆市高校市级口腔生物医学工程重点实验室
Author(s):
Yu Yong Tao Yu Deng Feng Wang Rui Chen Jun

Department of Orthodontics, Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Key Laboratory of Oral Biomedical Engineering of Chongqing Colleges and Universities, Stomatological Hospital of Chongqing Medical University, Chongqing, 401147, China

关键词:
人牙囊干细胞人牙髓干细胞体外共培养成骨分化
Keywords:
human dental follicle stem cells human dental pulp stem cells co-culture in vitro osteogenic differentiation
分类号:
R322.41;R329-33;R329.28
文献标志码:
A
摘要:

目的       通过体外建立人牙囊干细胞(human dental follicle stem cells, HDFSCs)和人牙髓干细胞(human dental pulp stem cells, HDPSCs)共培养体系,研究HDPSCs对HDFSCs增殖、成骨分化的作用。      方法      ①利用改良的组织块-消化联合法、免疫组化方法、流式细胞术以及多向诱导方法对HDFSCs和HDPSCs进行分离、培养、鉴定;②采用0.4 μm孔径的Transwell小室构建HDFSCs和HDPSCs体外共培养体系;③CCK-8测定HDFSCs增殖活性的变化;④与HDPSCs体外共培养1周,qRT-PCR检测HDFSCs几个标志性成骨基因:碱性磷酸酶(alkaline phosphatase, ALP)、人类相关转录因子2(runt-related transcription factor 2 , Runx2)、骨桥蛋白(osteopontin,OPN)、骨涎蛋白(bone sialoprotein,BSP)以及Ⅰ型胶原(collagen type 1,Col-1)的表达情况。      结果      ①成功分离、培养、鉴定HDFSCs和HDPSCs;②从第4天开始,共培养组HDFSCs的增殖活力明显高于对照组(P<0.05);③与对照组相比,共培养组ALP、Runx2、OPN、BSP、Col-1的相对表达量均明显增高(P<0.05)。      结论      与HDPSCs共培养能增强HDFSCs增殖活力及成骨分化。

Abstract:

Objective      To determine the role of human dental pulp stem cells (HDPSCs) in the proliferation and osteogenic differentiation of human dental follicle stem cells (HDFSCs) under co-culture in vitro.       Methods      ①HDFSCs and HDPSCs were isolated, cultured and identified by a modified tissue block digestion, immunohistochemistry, flow cytometry and multilineage inducement. ②Cell culture chamber(Transwell) with pore size of 0.4 μm in diameter were used to facilitate the co-culture system of HDFSCs and HDPSCs in vitro. ③CCK-8 assay was used to test the changes of proliferative activity of HDFSCs; ④ In 1 week after in vitro co-culture system of HDFSCs and HDPSCs, the expression of several osteoblast marker genes including the alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteopontin (OPN), bone sialoprotein (BSP) and collagen type 1 (Col-1) were measured with qRT-PCR.       Results      ①HDFSCs and HDPSCs were isolated, cultured and identified successfully. ②Starting from the fourth day, HDFSCs in the co-culture group showed a higher proliferative activity than that in the control group (P<0.05). ③Compared with the control group, there were significant increases in the relative mRNA levels of ALP, Runx2, OPN, BSP and Col-1 (P<0.05) in the co-culture group.       Conclusion      Co-culture with HDPSCs stimulates the proliferous activity and osteogenic differentiation in HDFSCs.

相似文献/References:

[1]王雪飞,张纲,裘松波,等.构建siRNA慢病毒载体调控人牙髓干细胞Delta1表达的实验研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(01):30.
 Wang Xuefei,Zhang Gang,Qiu Songbo,et al.Regulation of Delta1 expression in DPSCs by specific siRNA lentivirus vector[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(22):30.

更新日期/Last Update: 2015-11-21