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[比格犬牙髓干细胞的分离与鉴定]

[Isolation and identification of Beagle dog dental pulp stem cells].

作者信息

Wang Yuan-yuan, Zhao Yu-ming, Ge Li-hong

机构信息

Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology, Beijing 100081, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2012 Apr;47(4):241-5. doi: 10.3760/cma.j.issn.1002-0098.2012.04.012.

Abstract

OBJECTIVE

To identify the existence of the dental pulp stem cells in Beagle's pulp tissue by using the same methods of isolating and culturing the human dental pulp stem cells.

METHODS

Pulp tissue was extirpated from the crown and root of the Beagle's healthy permanent tooth, and digested by dispase for cell culture. Classical identification methods of mesenchymal stem cells including observation of biological characteristics, capacity of multilineage differentiation, and expression of specific markers associated with mesenchymal stem cells were applied to verify the existence of Beagle's dental pulp stem cells.

RESULTS

A clonogenic, rapidly proliferative population of cells were isolated from Beagle' pulp tissue. Under the same culture condition, the Beagle's dental pulp stem cells had a significant higher colony-forming unit-fibroblast (CFU-F) formation rate (150 colony/10(4) cells) than the dental pulp cells derived from the human pulp tissue (60 colony/10(4) cells). These cells also had the multilineage differentiation ability. They could be induced to form mineralized nodules, lipid droplets and chondrocytes. Furthermore these cells expressed the mesenchymal stem cell markers including STRO-1, CD146, alkaline phosphatase, nestin, vimentin and cytokeratin-18.

CONCLUSIONS

There are dental pulp stem cells in the Beagle's pulp tissue.

摘要

目的

采用与分离培养人牙髓干细胞相同的方法,鉴定比格犬牙髓组织中牙髓干细胞的存在。

方法

从比格犬健康恒牙的冠部和根部取出牙髓组织,用 dispase 消化进行细胞培养。应用间充质干细胞的经典鉴定方法,包括观察生物学特性、多向分化能力以及与间充质干细胞相关的特异性标志物的表达,来验证比格犬牙髓干细胞的存在。

结果

从比格犬牙髓组织中分离出一群具有克隆性、增殖迅速的细胞。在相同培养条件下,比格犬牙髓干细胞的集落形成单位 - 成纤维细胞(CFU - F)形成率(150 个集落 / 10⁴ 个细胞)显著高于来源于人牙髓组织的牙髓细胞(60 个集落 / 10⁴ 个细胞)。这些细胞还具有多向分化能力。它们可被诱导形成矿化结节、脂滴和软骨细胞。此外,这些细胞表达间充质干细胞标志物,包括 STRO - 1、CD146、碱性磷酸酶、巢蛋白、波形蛋白和细胞角蛋白 - 18。

结论

比格犬牙髓组织中存在牙髓干细胞。

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