Jin Mei, Yaling Yu, Zhibin Wang, Jianse Zhang
Anatomy Department & Institute of Bioscaffold Transplantation and Immunology, Wenzhou Medical University, 1210 Wenzhou University Town, Wenzhou, Zhejiang, 325035, China.
Methods Mol Biol. 2016;1397:53-63. doi: 10.1007/978-1-4939-3353-2_6.
The extracellular matrix (ECM) retains three-dimensional structures for the stimulation of cell growth, with components of the ECM relatively conserved between species. Interest in the use of decellularized scaffold-based strategies for organ regeneration is increasing rapidly. Decellularized scaffolds derived from animal organs are a promising material for organ engineering, with a number of prominent advances having been reported in the past few years.In this article we describe a simple and robust methodology for generating decellularized rat kidneys. To obtain these scaffolds, we perfuse rat kidneys with detergents through the abdominal aorta. After decellularization, kidney scaffolds are harvested for evaluation of vascular structure and histology. Qualitative evaluation involves vascular corrosion casting, transmission electron microscopy, and several different histological and immunofluorescent methods. SDS residue levels are assessed by ultraviolet-visible spectrophotometer (UV-VIS).
细胞外基质(ECM)保留三维结构以刺激细胞生长,其成分在物种间相对保守。对基于去细胞支架的器官再生策略的兴趣正在迅速增加。源自动物器官的去细胞支架是器官工程的一种有前途的材料,在过去几年中已有许多显著进展的报道。在本文中,我们描述了一种简单且可靠的制备去细胞大鼠肾脏的方法。为了获得这些支架,我们通过腹主动脉用去污剂灌注大鼠肾脏。去细胞后,收获肾脏支架以评估血管结构和组织学。定性评估包括血管铸型腐蚀、透射电子显微镜以及几种不同的组织学和免疫荧光方法。通过紫外可见分光光度计(UV-VIS)评估十二烷基硫酸钠残留水平。