Abstract
Many decellularized extracellular matrix-derived whole organs have been widely used in studies of tissue engineering and cancer models. However, decellularizing porcine esophagus to obtain decellularized esophageal matrix (DEM) for potential biomedical applications has not been widely investigated. In this study a modified decellularization protocol was employed to prepare a porcine esophageal DEM for the study of cancer cell growth. The cellular removal and retention of matrix components in the porcine DEM were fully characterized. The microstructure of the DEM was observed using scanning electronic microscopy. Human esophageal squamous cell carcinoma (ESCC) and human primary esophageal fibroblast cells (FBCs) were seeded in the DEM to observe their growth. Results show that the decellularization process did not cause significant loss of mechanical properties and that blood ducts and lymphatic vessels in the submucosa layer were also preserved. ESCC and FBCs grew on the DEM well and the matrix did not show any toxicity to cells. When FBS and ESCC were cocultured on the matrix, they secreted more periostin, a protein that supports cell adhesion on matrix. This study shows that the modified decellularization protocol can effectively remove the cell materials and maintain the microstructure of the porcine esophageal matrix, which has the potential application of studying cell growth and migration for esophageal cancer models.
Highlights
Esophageal cancer is the sixth leading cause of cancer-related deaths worldwide [1]with a very limited number of effective treatments
The purpose of this study was to determine the effectiveness of the decellularization process while maintaining the native microstructure of the extracellular matrix and to examine whether the decellularized matrix can support cell growth
The Pico-Green dsDNA assay showed that DNA removal through the decellularization protocol consistently removed approximately 92.5% of the quantifiable native porcine double-stranded DNA (n = 3) (p < 0.0001) (Figure 1I)
Summary
Esophageal cancer is the sixth leading cause of cancer-related deaths worldwide [1]with a very limited number of effective treatments. New tissue engineering strategies are being used to develop esophageal cancer models to replicate the tumor-specific cellular and matrix microenvironments Such efforts being made using tissue-engineering concepts are growing cancer cells in synthetic porous polymer scaffolds or nature collagen/Matrigel hydrogels to form artificial tumor tissue. These models have the potential to overcome the limitations of 2D monolayer culture [2,3,4,5], as they help mimic cancer growth, progression, and metastasis by providing closer matrix environments [6]. The rat esophagus does not contain submucosal glands in their esophagus, such as those found in pigs or humans [14], any attempt to recapitulate the human esophagus in the rat model would not contain all relevant anatomical attributes found in humans
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