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来自兔(穴兔)的牙龈间充质干细胞:分离、培养与鉴定

Gingival-Derived Mesenchymal Stem Cell from Rabbit (Oryctolagus cuniculus): Isolation, Culture, and Characterization.

作者信息

Nugraha Alexander Patera, Rantam Fedik Abdul, Narmada Ida Bagus, Ernawati Diah Savitri, Ihsan Igo Syaiful

机构信息

Department of Medical Science, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.

Department of Orthodontics, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.

出版信息

Eur J Dent. 2021 May;15(2):332-339. doi: 10.1055/s-0040-1719213. Epub 2020 Dec 1.

Abstract

OBJECTIVE

This study aims to confirm whether the GDMSCs isolated from rabbit's () gingiva are mesenchymal stem cells (MSCs).

MATERIALS AND METHODS

This study design was partly quasi-experimental with an observational design. GDMSCs were isolated from the gingiva of healthy male rabbits () ( = 2), 6 months old, and 3 to 5 kg of body weight. The specific cell surface markers of MSCs; clusters of differentiation (CD), namely, CD44, CD73, CD90, CD105, and CD200 expressions; and hematopoietic stem cell surface markers CD34 and CD45 were examined using flow cytometry and immunohistochemistry with immunofluorescence. The osteogenic differentiation of isolated GDMSCs was examined using alizarin red staining.

RESULTS

GDMSCs in the fourth passage showed a spindle-like formation and fibroblast-like cells that attached to the base of the culture plate. GDMSCs were MSCs that positively expressed CD44, CD73, CD90, CD105, and CD200 but did not express CD34 and CD45 when examined using flow cytometry and immunohistochemical analysis. GDMSCs had osteogenic differentiation confirmed by calcified deposits with a red-violet and brownish color after alizarin red staining.

CONCLUSION

GDMSCs isolated from the rabbits () were confirmed as MSCs documented using immunohistochemistry and flow cytometry. GDMSCs can differentiate into osteogenic lineage that may be suitable for regenerative dentistry.

摘要

目的

本研究旨在确认从兔()牙龈分离出的牙龈间充质干细胞(GDMSCs)是否为间充质干细胞(MSCs)。

材料与方法

本研究设计部分为准实验性观察性设计。从6月龄、体重3至5千克的健康雄性兔()( = 2)的牙龈中分离GDMSCs。使用流式细胞术和免疫荧光免疫组织化学检测MSCs的特异性细胞表面标志物;分化簇(CD),即CD44、CD73、CD90、CD105和CD200的表达;以及造血干细胞表面标志物CD34和CD45。使用茜素红染色检测分离的GDMSCs的成骨分化。

结果

第四代GDMSCs呈现纺锤样形态和成纤维细胞样细胞,附着于培养板底部。当使用流式细胞术和免疫组织化学分析检测时,GDMSCs是阳性表达CD44、CD73、CD90、CD105和CD200但不表达CD34和CD45的MSCs。茜素红染色后,GDMSCs有钙化沉积,呈红紫色和褐色,证实有成骨分化。

结论

从兔()分离出的GDMSCs经免疫组织化学和流式细胞术证实为MSCs。GDMSCs可分化为成骨谱系,可能适用于再生牙科。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f557/8184309/42de66623e90/10-1055-s-0040-1719213_9_0991_01.jpg

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