Decellularized Porcine Esophageal Scaffold Preparation: A Technique to Generate Acellular Scaffold from a Pig Esophagus
Transcript
Decellularized scaffolds derived from the extracellular matrix of specific tissue mimic their native extracellular microenvironment. These scaffolds retain their structural integrity and biochemical composition and serve as promising platforms for tissue engineering.
To prepare a decellularized porcine esophageal scaffold, begin with a freshly excised porcine esophagus. Bisect the esophagus longitudinally and rinse it with a suitable buffer to remove any debris. Next, secure the esophagus with its mucosal surface, the innermost layer, facing up.
Separate the mucosa from the underlying submucosa at one end of the esophagus. Dissect the esophagus longitudinally along the loose connective tissue of the mucosa called the lamina propria to fully separate the mucosal surface from the underlying submucosa.
Further, excise the proximal and distal regions of the mucosa, which contain a higher density of submucosal glands and a thicker lamina propria layer. Cut the mucosal tissue into uniformly sized pieces.
Treat the tissue with a hypertonic saline solution supplemented with antibiotics. Antibiotics help prevent bacterial growth. At high solute concentration, water moves out from the cells and causes their shrinkage. This step further aids in cellular death and detachment from the extracellular matrix.
Now, gently detach the loosened epithelium from the underlying tissue. Rinse the tissue with buffer to remove cellular debris. Finally, incubate the tissue in glycerol, which further facilitates tissue decellularization by dehydrating and lysing cells while preserving the integrity of the basement membrane.
