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人脐带血来源无限制体干细胞在 3Dβ-磷酸三钙-藻酸盐-明胶支架上的软骨分化。

Chondrogenic Differentiation of Human Umbilical Cord Blood-Derived Unrestricted Somatic Stem Cells on A 3D Beta-Tricalcium Phosphate-Alginate-Gelatin Scaffold.

机构信息

Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Cellular and Molecular Research Centre, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

出版信息

Cell J. 2014 Feb 3;16(1):43-52.

Abstract

OBJECTIVE

Finding cell sources for cartilage tissue engineering is a critical procedure. The purpose of the present experimental study was to test the in vitro efficacy of the beta-tricalcium phosphate-alginate-gelatin (BTAG) scaffold to induce chondrogenic differentiation of human umbilical cord blood-derived unrestricted somatic stem cells (USSCs).

MATERIALS AND METHODS

In this experimental study, USSCs were encapsulated in BTAG scaffold and cultured for 3 weeks in chondrogenic medium as chondrogenic group and in Dulbecco's Modified Eagle's Medium (DMEM) as control group. Chondrogenic differentiation was evaluated by histology, immunofluorescence and RNA analyses for the expression of cartilage extracellular matrix components. The obtain data were analyzed using SPSS version 15.

RESULTS

Histological and immunohistochemical staining revealed that collagen II was markedly expressed in the extracellular matrix of the seeded cells on scaffold in presence of chondrogenic media after 21 days. Reverse transcription-polymerase chain reaction (RT-PCR) showed a significant increase in expression levels of genes encoded the cartilage-specific markers, aggrecan, type I and II collagen, and bone morphogenetic protein (BMP)-6 in chondrogenic group.

CONCLUSION

This study demonstrates that BTAG can be considered as a suitable scaffold for encapsulation and chondrogenesis of USSCs.

摘要

目的

寻找用于软骨组织工程的细胞来源是一个关键步骤。本实验研究的目的是测试β-磷酸三钙-藻酸盐-明胶(BTAG)支架在体外诱导人脐带血来源的无限制体干细胞(USSCs)向软骨分化的功效。

材料与方法

在这项实验研究中,USSCs 被包裹在 BTAG 支架中,并在软骨形成培养基中培养 3 周作为软骨形成组,在 DMEM 中作为对照组。通过组织学、免疫荧光和 RNA 分析评估软骨细胞外基质成分的表达来评估软骨分化。使用 SPSS 版本 15 对获得的数据进行分析。

结果

组织学和免疫组织化学染色显示,在存在软骨形成培养基 21 天后,细胞外基质中明显表达了胶原蛋白 II。逆转录-聚合酶链反应(RT-PCR)显示,软骨形成组中编码软骨特异性标志物的基因,聚集蛋白聚糖、I 型和 II 型胶原以及骨形态发生蛋白(BMP)-6 的表达水平显著增加。

结论

本研究表明,BTAG 可被视为 USSCs 包封和软骨形成的合适支架。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3345/3933438/80b70c31c947/Cell-J-16-43-g01.jpg

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