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一种新的间隔穿孔修复方法:间隔软骨细胞在体外诱导 hASC 软骨形成。

A Novel Approach to Septal Perforation Repair: Septal Cartilage Cells Induce Chondrogenesis of hASCs In Vitro.

机构信息

Faculty of Engineering and Architecture, Department of Genetics and Bioengineering, Yeditepe University, Istanbul, Turkey.

出版信息

Appl Biochem Biotechnol. 2019 Aug;188(4):942-951. doi: 10.1007/s12010-019-02964-9. Epub 2019 Feb 11.

Abstract

The aim of this study was to investigate the effect of medium harvested from septal cartilage cells on chondrogenic differentiation of adipose stem cells (hASCs) and to compare/contrast its properties to those of a commonly used standard medium formulation in terms of induction and maintenance of chondrogenic hASCs. Differentiation was carried out under three different conditions: septal cartilage medium-SCM, chondrogenic differentiation medium-CM, and 50:50 mixture of CM/SCM. Mesenchymal stem cells (MSCs) markers were determined by flow cytometry. The cytotoxic and apoptotic effects were determined by MTS and Annexin V assay, respectively. The differentiation status of the cells was confirmed by Alcian blue staining, and quantitative real-time flow cytometry showed that hASCs were positive for MSCs, negative for hematopoietic stem cells and endothelial cell surface markers. According to MTS analysis, the first condition was not toxic at any concentration tested. Annexin V assay revealed that the application of different concentrations of SCM did not result in any cell death. The Alcian blue and gene expression analyses showed that the cells in the SCM group underwent the highest cartilage cell formation. The observed increase in chondrogenesis may offer better treatment options for the cartilage defects seen in nasal septum perforation.

摘要

本研究旨在探讨鼻中隔软骨细胞培养液对脂肪干细胞(hASC)软骨分化的影响,并比较/对比其特性与常用标准培养基配方在诱导和维持软骨 hASC 方面的特性。分化在三种不同条件下进行:鼻中隔软骨培养基-SCM、软骨分化培养基-CM 和 CM/SCM 混合物的 50:50 混合物。通过流式细胞术测定间充质干细胞标志物。通过 MTS 和 Annexin V 测定分别确定细胞的细胞毒性和凋亡作用。通过阿尔辛蓝染色确认细胞的分化状态,实时定量流式细胞术显示 hASC 对间充质干细胞呈阳性,对造血干细胞和内皮细胞表面标志物呈阴性。根据 MTS 分析,任何测试浓度的第一种条件都没有毒性。Annexin V 测定显示,不同浓度 SCM 的应用不会导致任何细胞死亡。阿尔辛蓝和基因表达分析表明,SCM 组的细胞经历了最高的软骨细胞形成。观察到的软骨形成增加可能为鼻中隔穿孔中所见的软骨缺陷提供更好的治疗选择。

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