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体外诱导脂肪间充质干细胞向软骨细胞分化及其与关节软骨细胞表型比较。

Comparison between Chondrogenic Markers of Differentiated Chondrocytes from Adipose Derived Stem Cells and Articular Chondrocytes In Vitro.

机构信息

Department of Anatomical Sciences and Molecular Biology, Medical School, Esfahan University of Medical Sciences, Esfahan, Islamic Republic of Iran.

出版信息

Iran J Basic Med Sci. 2013 Jun;16(6):763-73.

Abstract

OBJECTIVE(S): Osteoarthritis is one of the most common diseases in middle-aged population in the world. Cartilage tissue engineering (TE) has been presented as an effort to introduce the best combination of cells, biomaterial scaffolds and stimulating growth factors to produce a cartilage tissue similar to the natural articular cartilage. In this study, the chondrogenic potential of adipose derived stem cells (ADSCs) was compared with natural articular chondrocytes cultured in alginate scaffold.

MATERIALS AND METHODS

Human ADSCs were obtained from subcutaneous adipose tissue and human articular chondrocytes from non-weight bearing areas of knee joints. Cells were seeded in 1.5% alginate and cultured in chondrogenic media for three weeks with and without TGFβ3. The genes expression of types II and X collagens was assessed by Real Time PCR and the amount of aggrecan (AGC) and type I collagen measured by ELISA and the content of glycosaminoglycan evaluated by GAG assay.

RESULTS

Our findings showed that type II collagen, GAG and AGC were expressed, in differentiated ADSCs. Meanwhile, they produced a lesser amount of types II and X collagens but more AGC, GAG and type I collagen in comparison with natural chondrocytes (NCs).

CONCLUSION

Further attempt should be carried out to optimize achieving type II collagen in DCs, as much as, natural articular chondrocytes and decline of the production of type I collagen in order to provide efficient hyaline cartilage after chondrogenic induction, prior to the usage of harvested tissues in clinical trials.

摘要

目的

骨关节炎是世界中年人群体中最常见的疾病之一。软骨组织工程(TE)旨在引入最佳的细胞、生物材料支架和刺激生长因子组合,以产生类似于天然关节软骨的软骨组织。在这项研究中,比较了脂肪来源干细胞(ADSCs)的软骨生成潜力与在藻酸盐支架中培养的天然关节软骨细胞。

材料和方法

从皮下脂肪组织中获得人 ADSC,从膝关节非承重区域获得人关节软骨细胞。细胞接种在 1.5%的藻酸盐中,并在软骨形成培养基中培养 3 周,有和没有 TGFβ3。通过实时 PCR 评估 II 型和 X 型胶原的基因表达,通过 ELISA 测量聚集蛋白聚糖(AGC)和 I 型胶原的量,并通过 GAG 测定评估糖胺聚糖的含量。

结果

我们的研究结果表明,分化的 ADSC 中表达了 II 型胶原、GAG 和 AGC。同时,与天然软骨细胞(NCs)相比,它们产生的 II 型和 X 型胶原较少,但 AGC、GAG 和 I 型胶原较多。

结论

应该进一步尝试优化 DCs 中 II 型胶原的产生,尽可能与天然关节软骨细胞相同,并减少 I 型胶原的产生,以便在临床试验中使用采集的组织之前,在软骨诱导后提供高效的透明软骨。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b977/3758031/fc1ab9ac5415/ijbms-16-763-g001.jpg

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