天然支架对人脂肪源性间充质干细胞成软骨分化能力的评估

Mohsen Sheykhhasan, M. Ghiasi, Hossien Bakhtiyari Pak
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引用次数: 4

摘要

由于缺乏血管和低细胞密度,软骨修复损伤的能力受到限制。最近,作为解决这一问题的一种方法,组织工程明显优于其他治疗方法。无论细胞来源如何,组织工程的关键因素之一是选择合适的支架,这对于体内和体外组织的愈合和更新过程至关重要。利用胶原酶从吸脂术中分离脂肪组织间充质干细胞。流式细胞术验证后,将脂肪源性间充质干细胞(ADMSCs)分别包埋在海藻酸盐和琼脂糖支架中;然后在软骨培养基中培养3周。采用MTT法和组织学分析评价海藻酸盐和琼脂糖支架的功能。此外,采用Real-time PCR分析软骨形成基因的表达。所得数据采用SPSS软件进行统计学分析。海藻酸盐组和琼脂糖组在维持细胞活力方面存在显著差异,但在实时荧光定量PCR分析成软骨分化时,统计分析显示细胞/海藻酸盐组、细胞/琼脂糖组和间充质干细胞中聚集蛋白(aggrecan,作为软骨细胞特异性基因)和胶原II (collagen II,作为软骨细胞特异性基因)的表达差异显著(p<0.05)。组织学分析证实细胞有软骨细胞分化。藻酸盐支架可为脂肪源间充质干细胞的软骨分化提供适宜的环境。关键词:海藻酸;琼脂糖;Chondrogenic分化;间充质干细胞;组织工程
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The assessment of natural scaffolds ability in chondrogenic differentiation of human adipose-derived mesenchymal stem cells
The ability of cartilage to repair damage is limited due to lack of blood vessels and low cell density. Recently, tissue engineering is considerably preferred to other treatments as a way to solve this problem. Regardless of cell sources, one of the crucial factors in tissue engineering is to select an appropriate scaffold, which is essential for healing and renewal procedure of tissues in vivo and in vitro . Mesenchymal stem cells (MSCs) were isolated from adipose tissue in liposuction surgeries by use of collagenase enzyme. After verification by flow cytometry methods, adipose-derived mesenchymal stem cells (ADMSCs) were embedded into alginate and agarose scaffolds, separately; and then they were cultured in chondrogenic medium for 3 weeks. The ability of alginate and agarose scaffolds was assessed by use of MTT assay and histological analysis. In addition, analysis of chondrogenic genes expression by Real-time PCR was done. The obtained data were analyzed statistically by means of SPSS software. There was a significant difference between alginate and agarose groups in maintaining cells viable but, about chondrogenic differentiation analyzed by use of real-time PCR, statistical analysis has shown a significant difference in expression of aggrecan (as a chondrocyte-specific gene) and collagen II (as an chondrocyte-specific gene) between cell/alginate and cell/agarose and MSCs (p<0.05). Chondrocyte differentiation of cells was verified by histological analysis. Alginate scaffold can provide a suitable environment for chondrogenic differentiation of  adipose derived mesenchymal stem cells. Keywords: Alginate; Agarose; Chondrogenic differentiation; Mesenchymal stem cell; Tissue engineering
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