Yu Yaling, Liu Li, Zhang Jianse, Wei Zairong, Mei Jin
Department of Orthopedic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.
Department of Plastic Surgery, Affiliated Hospital of Zunyi Medical College, Zunyi, 563000, China.
Methods Mol Biol. 2018;1577:111-119. doi: 10.1007/7651_2017_72.
The decellularized (DC) scaffolds retain three-dimensional structures for the stimulation of cell growth, with components of the extracellular matrix (ECM) relatively conserved between species. The strategy based on decellularized scaffolds provides a new way for organ regeneration, with a number of prominent advances having been reported in the past few years. While their lack of biomechanical strength and excessive degradation limit the clinical applications, therefore it is urgent to modify the DC scaffolds to improve the performance.In this article we describe a simple and robust modification protocol for DC rat kidney scaffolds. To modify, we perfuse DC rat kidneys with glutaraldehyde through the perfusion circulation of the decellularization. After cross-linking, kidney scaffolds are harvested for evaluation of histology, structural stability, and biocompatibility, involving water absorption testing, biomechanical testing, scanning electron microscopy, and several different histological and immunofluorescent analyses.
去细胞(DC)支架保留了用于刺激细胞生长的三维结构,细胞外基质(ECM)的成分在物种间相对保守。基于去细胞支架的策略为器官再生提供了一条新途径,在过去几年中已有多项显著进展被报道。然而,它们缺乏生物力学强度和过度降解限制了其临床应用,因此迫切需要对DC支架进行改性以提高其性能。在本文中,我们描述了一种用于DC大鼠肾脏支架的简单且可靠的改性方案。为了进行改性,我们通过去细胞过程的灌注循环用戊二醛灌注DC大鼠肾脏。交联后,收获肾脏支架以评估组织学、结构稳定性和生物相容性,包括吸水性测试、生物力学测试、扫描电子显微镜以及几种不同的组织学和免疫荧光分析。