Abstract
Tissue microenvironments are rich in signaling molecules. However, factors in the tissue matrix that can serve as tissue-specific cues for engineering pancreatic tissues have not been thoroughly identified. In this study, we performed a comprehensive proteomic analysis of porcine decellularized pancreatic extracellular matrix (dpECM). By profiling dpECM collected from subjects of different ages and genders, we showed that the detergent-free decellularization method developed in this study permits the preservation of approximately 62.4% more proteins than a detergent-based method. In addition, we demonstrated that dpECM prepared from young pigs contained approximately 68.5% more extracellular matrix proteins than those prepared from adult pigs. Furthermore, we categorized dpECM proteins by biological process, molecular function, and cellular component through gene ontology analysis. Our study results also suggested that the protein composition of dpECM is significantly different between male and female animals while a KEGG enrichment pathway analysis revealed that dpECM protein profiling varies significantly depending on age. This study provides the proteome of pancreatic decellularized ECM in different animal ages and genders, which will help identify the bioactive molecules that are pivotal in creating tissue-specific cues for engineering tissues in vitro.
Highlights
Tissue matrix is a natural, bioactive material that provides unique microenvironments for tissue and organ formation and function
We have recently reported that decellularized pancreatic extracellular matrix prepared from rat pancreata can enhance human pancreatic tissue development during human pluripotent stem cell pancreatic differentiation with both induced pluripotent stem cells and embryonic stem cells
We demonstrated the advantage of the non-detergent treatment to preserve more porcine tissue matrix proteins during decellularization
Summary
Tissue matrix is a natural, bioactive material that provides unique microenvironments for tissue and organ formation and function. Such postnatal acquisition of functional islets hints at unique signals in newborn tissue/organ microenvironments [14,17] These studies imply that dpECM prepared from young animals may contain strong instructive cues such as unique proteins serving as the signaling molecules that can promote the development of pancreatic tissues. We used liquid chromatography-tandem mass spectrometry (LC-MS/MS), a more advanced LC-MS method, to quantify protein expression of the dpECM as opposed to other similar bottom-up approaches This technology directly searches against large existing databases to identify protein sequences immediately, unlike non-LC–MS-based mass spectrometry methods like matrix-assisted laser desorption and ionization (MALDI) and surface-enhanced laser desorption and ionization (SELDI) [31,32]. LC–MS/MS is more sensitive and less labor intensive than standard LC–MS, with higher retrieval rates of hydrophobic proteins than gel-analysis based MS methods, making it ideal for analysis of dpECM [31,32,33]
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