Solchaga Luis A, Penick Kitsie J, Welter Jean F
Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, USA.
Methods Mol Biol. 2011;698:253-78. doi: 10.1007/978-1-60761-999-4_20.
It is well known that adult cartilage lacks the ability to repair itself; this makes articular cartilage a very attractive target for tissue engineering. The majority of articular cartilage repair models attempt to deliver or recruit reparative cells to the site of injury. A number of efforts are directed to the characterization of progenitor cells and the understanding of the mechanisms involved in their chondrogenic differentiation. Our laboratory has focused on cartilage repair using mesenchymal stem cells and studied their differentiation into cartilage. Mesenchymal stem cells are attractive candidates for cartilage repair due to their osteogenic and chondrogenic potential, ease of harvest, and ease of expansion in culture. However, the need for chondrogenic differentiation is superposed on other technical issues associated with cartilage repair; this adds a level of complexity over using mature chondrocytes. This chapter will focus on the methods involved in the isolation and expansion of human mesenchymal stem cells, their differentiation along the chondrogenic lineage, and the qualitative and quantitative assessment of chondrogenic differentiation.
众所周知,成年软骨缺乏自我修复能力;这使得关节软骨成为组织工程极具吸引力的目标。大多数关节软骨修复模型试图将修复细胞递送至损伤部位或招募修复细胞至损伤部位。许多努力都致力于祖细胞的特性表征以及对其软骨形成分化所涉及机制的理解。我们实验室专注于使用间充质干细胞进行软骨修复,并研究它们向软骨的分化。间充质干细胞因其成骨和成软骨潜力、易于获取以及易于在培养中扩增,是软骨修复极具吸引力的候选细胞。然而,软骨形成分化的需求叠加在与软骨修复相关的其他技术问题上;这比使用成熟软骨细胞增加了一层复杂性。本章将重点介绍人骨髓间充质干细胞的分离和扩增方法、它们沿软骨形成谱系的分化以及软骨形成分化的定性和定量评估。