Role of in vitro co-culture of human dental pulp stem cells in proliferation and osteogenic differentiation of human dental follicle stem cells

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Role of in vitro co-culture of human dental pulp stem cells in proliferation and osteogenic differentiation of human dental follicle stem cells

更新时间：2026-01-09

- Role of in vitro co-culture of human dental pulp stem cells in proliferation and osteogenic differentiation of human dental follicle stem cells
- Journal of Army Medical University Vol. 37, Issue 22, Pages: 2267-2272(2015)
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  CLC： R322.41;R329-33;R329.28
- Published：2015
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余勇,陶昱,邓锋,王睿,陈军.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,Wang Rui,Chen Jun.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.
余勇,陶昱,邓锋,王睿,陈军.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. DOI：

　Yu Yong,Tao Yu,Deng Feng,Wang Rui,Chen Jun.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. DOI：

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（P0.05）；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（P0.05）。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 (P0.05). ③Compared with the control group

there were significant increases in the relative mRNA levels of ALP

Runx2

OPN

BSP and Col-1 (P0.05) in the co-culture group. Conclusion Co-culture with HDPSCs stimulates the proliferous activity and osteogenic differentiation in HDFSCs.

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Related Institution

Department of Periodontology， The Affiliated Stomatological Hospital of Chongqing Medical University

Chongqing Key Laboratory of Oral Diseases

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