Youngstrom Daniel W, Barrett Jennifer G
Department of Large Animal Clinical Sciences, Marion duPont Scott Equine Medical Center, Virginia Tech, Leesburg, VA, USA.
Methods Mol Biol. 2016;1502:195-202. doi: 10.1007/7651_2016_332.
Tendon bioreactors combine cells, scaffold, and mechanical stimulation to drive tissue neogenesis ex vivo. Faithful recapitulation of the native tendon microenvironment is essential for stimulating graft maturation or modeling tendon biology. As the mediator between cells and mechanical stimulation, the properties of a scaffold constitute perhaps the most essential elements in a bioreactor system. One method of achieving native scaffold properties is to process tendon allograft in a manner that removes cells without modifying structure and function: "decellularization." This chapter describes (1) production of tendon scaffolds derived from native extracellular matrix, (2) preparation of cell-laden scaffolds prior to bioreactor culture, and (3) tissue processing post-harvest for gene expression analysis. These methods may be applied for a variety of applications including graft production, cell priming prior to transplantation and basic investigations of tendon cell biology.
肌腱生物反应器将细胞、支架和机械刺激相结合，以在体外驱动组织新生。忠实重现天然肌腱微环境对于促进移植物成熟或模拟肌腱生物学至关重要。作为细胞与机械刺激之间的介质，支架的特性可能是生物反应器系统中最关键的要素。实现天然支架特性的一种方法是以去除细胞但不改变结构和功能的方式处理同种异体肌腱移植物：“去细胞化”。本章介绍了（1）源自天然细胞外基质的肌腱支架的制备，（2）生物反应器培养前负载细胞的支架的制备，以及（3）收获后用于基因表达分析的组织处理。这些方法可应用于多种用途，包括移植物生产、移植前细胞预处理以及肌腱细胞生物学的基础研究。
