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人诱导多能干细胞向软骨细胞的分化

Differentiation of Human Induced Pluripotent Stem Cells to Chondrocytes.

作者信息

Guzzo Rosa M, Drissi Hicham

机构信息

Department of Orthopaedic Surgery, MC4037, UConn Health, 263 Farmington Avenue, Farmington, CT, 06030, USA.

Stem Cell Institute, UConn Health, Farmington, CT, USA.

出版信息

Methods Mol Biol. 2015;1340:79-95. doi: 10.1007/978-1-4939-2938-2_6.

Abstract

Human induced pluripotent stem (iPS) cells are relevant tools for modeling human skeletal development and disease, and represent a promising source of patient-specific cells for the regeneration of skeletal tissue, such as articular cartilage. Devising efficient and reproducible strategies, which closely mimic the physiological chondrogenic differentiation process, will be necessary to generate functional chondrocytes from human iPS cells. Our previous study demonstrated the generation of chondrogenically committed human iPS cells via the enrichment of a mesenchymal-like progenitor population, application of appropriate high-density culture conditions, and stimulation with bone morphogenetic protein-2 (Bmp-2). The differentiated iPS cells showed temporal expression of cartilage genes and the accumulation of a cartilaginous extracellular matrix in vitro. In this chapter, we provide detailed methodologies for the differentiation of human iPS cells to the chondrogenic lineage and describe protocols for the analysis of chondrogenic differentiation.

摘要

人诱导多能干细胞(iPS细胞)是用于模拟人类骨骼发育和疾病的相关工具,并且是用于骨骼组织(如关节软骨)再生的有前景的患者特异性细胞来源。设计有效且可重复的策略,紧密模拟生理性软骨形成分化过程,对于从人iPS细胞生成功能性软骨细胞是必要的。我们之前的研究表明,通过富集间充质样祖细胞群体、应用适当的高密度培养条件以及用骨形态发生蛋白-2(Bmp-2)刺激,可生成软骨定向分化的人iPS细胞。分化的iPS细胞在体外显示出软骨基因的时序表达以及软骨细胞外基质的积累。在本章中,我们提供了将人iPS细胞分化为软骨谱系的详细方法,并描述了软骨分化分析的方案。

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