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人出生时牙髓干细胞向胰岛素生成细胞的分化潜能

The Differentiation Potential of Human Natal Dental Pulp Stem Cells into Insulin-Producing Cells.

作者信息

Suchanek J, Nasry S A, Soukup T

机构信息

Department of Dentistry, Charles University - Faculty of Medicine in Hradec Králové, Hradec Králové, Czech Republic.

Department of Surgery and Oral Medicine, Oro-dental division, National Research Centre, Cairo, Egypt.

出版信息

Folia Biol (Praha). 2017;63(4):132-138. doi: 10.14712/fb2017063040132.

Abstract

Mesenchymal stem cells have the ability to differentiate into insulin-producing cells, raising the hope for diabetes mellitus treatment. The aim of this research was to study the ability of stem cells from discarded natal teeth to differentiate into insulinproducing cells. Two vital human natal teeth were obtained from a healthy 2-day-old female. Stem cells from the dental pulp were isolated, cultured under xenogenic-free conditions, propagated and characterized. Proliferative activity, population doubling time and viability were measured, and the multipotent differentiation ability was investigated. A twostep protocol was used to induce the human natal dental pulp stem cells to differentiate into insulinproducing cells. Phenotypic analysis was done using flow cytometry. Immunohistochemistry was performed to detect insulin and C-peptide. PDX1, HES1 and Glut2 gene expression analysis was performed by quantitative reverse transcription-polymerase chain reaction. Human natal dental pulp stem cells were able to undergo osteogenic, chondrogenic and adipogenic differentiation upon exposure to the specific differentiation media for each lineage. Their differentiation into insulin-producing cells was confirmed by expression of C-peptide and insulin, as well as by 975.4 % higher expression of PDX-1 and 469.5 % higher expression of HES1 in comparison to the cells cultivated in standard cultivation media. Glut2 transporter mRNA was absent in the non-differentiated cells, and differentiation of the stem cells into insulin-producing cells induced appearance of the mRNA of this transporter. We were the first to demonstrate that stem cells obtained from the pulp of natal teeth could be differentiated into insulinproducing cells, which might prove useful in the stem cell therapy for type 1 diabetes.

摘要

间充质干细胞具有分化为胰岛素生成细胞的能力,这为糖尿病治疗带来了希望。本研究的目的是研究废弃乳牙来源的干细胞分化为胰岛素生成细胞的能力。从一名健康的2日龄女性获取了两颗新鲜的乳牙。分离牙髓干细胞,在无外源条件下培养、传代并进行鉴定。测定增殖活性、群体倍增时间和活力,并研究多能分化能力。采用两步方案诱导人乳牙牙髓干细胞分化为胰岛素生成细胞。使用流式细胞术进行表型分析。进行免疫组织化学检测胰岛素和C肽。通过定量逆转录-聚合酶链反应进行PDX1、HES1和Glut2基因表达分析。人乳牙牙髓干细胞在暴露于各谱系的特定分化培养基后能够进行成骨、软骨和成脂分化。与在标准培养基中培养的细胞相比,它们分化为胰岛素生成细胞通过C肽和胰岛素的表达得以证实,同时PDX-1表达升高975.4%,HES1表达升高469.5%。未分化细胞中不存在Glut2转运蛋白mRNA,而干细胞分化为胰岛素生成细胞诱导了该转运蛋白mRNA的出现。我们首次证明从乳牙牙髓获得的干细胞可以分化为胰岛素生成细胞,这可能对1型糖尿病的干细胞治疗有用。

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