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体外用人胚牙细胞条件培养液诱导脐带间充质干细胞向成牙本质样细胞分化。

Differentiation of mesenchymal stem cells from human umbilical cord tissue into odontoblast-like cells using the conditioned medium of tooth germ cells in vitro.

机构信息

Department of Prosthodontics, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, China.

出版信息

Biomed Res Int. 2013;2013:218543. doi: 10.1155/2013/218543. Epub 2013 May 13.

Abstract

The easily accessible mesenchymal stem cells in the Wharton's jelly of human umbilical cord tissue (hUCMSCs) have excellent proliferation and differentiation potential, but it remains unclear whether hUCMSCs can differentiate into odontoblasts. In this study, mesenchymal stem cells were isolated from the Wharton's jelly of human umbilical cord tissue using the simple method of tissue blocks culture attachment. UCMSC surface marker expression was then evaluated for the isolated cells using flow cytometry. The third-passage hUCMSCs induced by conditioned medium from developing tooth germ cells (TGC-CM) displayed high alkaline phosphatase (ALP) levels (P < 0.001), an enhanced ability to proliferate (P < 0.05), and the presence of mineralized nodules. These effects were not observed in cells treated with regular medium. After induction of hUCMSCs, the results of reverse transcriptional polymerase chain reaction (PCR) indicated that the dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) genes were significantly tested. Additionally, dentin sialoprotein (DSP) and DMP1 demonstrated significant levels of staining in an immunofluorescence analysis. In contrast, the control cells failed to display the characteristics of odontoblasts. Taken together, these results suggest that hUCMSCs can be induced to differentiate into odontoblast-like cells with TGC-CM and provide a novel strategy for tooth regeneration research.

摘要

人脐带组织华通氏胶中的间充质干细胞(hUCMSCs)易于获取,具有极好的增殖和分化潜能,但 hUCMSCs 是否能分化为成牙本质细胞仍不清楚。本研究采用组织块培养贴壁的简单方法,从人脐带组织华通氏胶中分离间充质干细胞,然后用流式细胞术评估分离细胞的 UCMSC 表面标志物表达。用牙胚细胞条件培养基(TGC-CM)诱导的第 3 代 hUCMSCs 碱性磷酸酶(ALP)水平显著升高(P<0.001),增殖能力增强(P<0.05),并形成矿化结节。用常规培养基处理的细胞则未观察到这些效应。hUCMSCs 诱导后,反转录聚合酶链反应(PCR)的结果表明,牙本质涎磷蛋白(DSPP)和牙本质基质蛋白 1(DMP1)基因得到了显著检测。此外,免疫荧光分析显示牙本质涎蛋白(DSP)和 DMP1 有明显染色。相比之下,对照细胞未能显示出牙本质细胞的特征。综上所述,这些结果表明,hUCMSCs 可以在 TGC-CM 的诱导下分化为成牙本质细胞样细胞,为牙齿再生研究提供了新策略。

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