Decellularization of Rat Kidneys to Produce Extracellular Matrix Scaffolds

Methods Mol Biol. 2016:1397:53-63. doi: 10.1007/978-1-4939-3353-2_6.

Abstract

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).

Keywords: Decellularized scaffolds; Extracellular matrix; Kidney; Regeneration; Tissue/organ engineering.

MeSH terms

  • Animals
  • Extracellular Matrix / metabolism*
  • Fluorescent Antibody Technique
  • Immunohistochemistry
  • Kidney* / cytology
  • Kidney* / metabolism
  • Kidney* / ultrastructure
  • Rats
  • Tissue Scaffolds*