Project description:The extracellular matrix is an important part of the cellular microenvironment regulating various cellular functions. Decellularized matrices present a valuable strategy for recreating the cellular niche in vitro and implementing matrix signaling in cell culture models. However, the tissue-specificity of decellularized matrices is little considered in most in vitro models. Here, we present the generation of a porcine-derived pancreas-specific decellularized extracellular matrix scaffold (d-PanMa) and show the impact of scaffold-specific cues on stem cell culture. The proteomic analysis in this work compare the proteome of porcine pancreas (PanMa), intestine (SISser) and lung (lungMa) both in its native and decellularized form. The analysis verify the enrichment of important matrisome proteins in the decellular matrices and provide a detailed map of the protein composition and abundance across the matrices.
Project description:Porcine cytomegalovirus (PCMV; genus Cytomegalovirus, subfamily Betaherpesvirinae, family Herpesviridae) is an immunosuppressive virus that mainly inhibits the immune function of T lymphocytes and macrophages, which has caused great distress to the farming industry. In this study, we obtained the miRNA expression profiles of PCMV-infected and control porcine macrophages, PCMV-infected and control porcine tissues via high-throughput sequencing. The comprehensive analysis of miRNA profiles showed that 306 miRNA database annotated and 295 novel pig-encoded miRNAs were detected. Gene Ontology (GO) analysis of the target genes of miRNAs in PCMV infected porcine macrophages showed that the differentially expressed miRNAs are mainly involved in immune and metabolic process. This is the first report of the miRNA transcriptome in PCMV infected porcine macrophages and PCMV infected tissues and the analysis of the miRNA regulatory mechanism during PCMV infection. Further research into the regulatory mechanisms of miRNAs during immunosuppressive viral infections will contribute to the treatment and prevention of immunosuppressive viruses. miRNA expression profiling of PCMV-infected and control porcine macrophages; PCMV-infected and control porcine tissues via high-throughput sequencing.
Project description:In this study we have employed RNA seq on ten different tissues including four brain tissues from two boars to gain a understanding of the differential variations in transcriptional profiles for these tissues consisting of occipital cortex, frontal cortex, hypothalamus and cerebellum along with such diverse tissues as heart, spleen, liver, kidney, lung and musculus longissimus dorsi. This has enabled us to perform comparative gene expression analysis of brain regions versus non-brain tissues along with inter-brain tissue comparisons. Hence, we have tested for differentially expressed genes and isoforms, differential splicing, transcription start sites (TSS), and differential promoter usage between all ten porcine tissues.
Project description:Decellularized extracellular matrix (dECM)-based hydrogels combined with hASCs could serve as an interface for studying behavior and differentiation properties in a cartilage microenvironment. In the present study, we described the behavior of hASCs cultured in a commercial dECM MatriXpec™. The protein composition of MatriXpec™ was accessed by mass spectrometry and is mainly represented by collagen proteins, building an hydrogel fibrous ultrastructure.
Project description:Hitherto, microenvironments provided by dermal analogues could not avoid frequent split-thickness skin grafts (STSGs) failure and scar fibrosis. We reported a novel method assembling 3D-printed dermal analogues (PDAs) with porcine dermal extracellular matrix(dECM)'s recapitulated the latter’s compositional and topological features. These optimized PADs reduced skin wounds contraction and improved cosmetic upshots proving superior to STSGs and commercially available dermal templates. Once grafted onto wounds beds, optimized PDAs evoked a type-2-like immune response, activated type-2 macrophages (M2-Mφ), upregulated anti-inflammatory genes like Wnt11, Fgf16 and ATF3, downregulated profibrotic genes like Il13r⍺2, Itg⍺11, and IL1β, and hindered fibrosis. Thus, PDAs' beneficial effects resulted from preserved dermis-specific ECM cues (by mass spectrometry analysis we identified more than 200 unique protein species inside our dECM powder) and well-balanced physicochemical properties, which collaboratively modulated crucial endogenous signaling pathways.
Project description:comparison of pooled decellularized porcine left ventricle (LV) or sinoatrial node (SAN) extracellular matrix digested with either trypsin or elastase
Project description:To expand the knowledge on the porcine salivary proteome, secretions from the three major salivary glands were collected from anaesthetised piglets. Pilocarpine and isoproterenol were simultaneously injected intraperitoneally to increase the volume and protein concentration of the saliva, respectively. The protein composition and relative protein-specific abundance of saliva secreted by the parotid gland and by the mandibular and monostomatic sublingual gland, were determined using iTRAQ. When combining two detection methods, MALDI-TOF/TOF MS and Q-Exactive orbitrap MS/MS, a total of 122 porcine salivary proteins and 6 mammalian proteins with a predicted porcine homolog were identified. Only a quantitative and not a qualitative difference was observed between both ductal secretions. The 128 proteins were detected in both secretions, however, at different levels. Twenty-four proteins (20 porcine, 4 mammalian with a predicted porcine homolog) were predominantly secreted by the parotid gland, such as carbonic anhydrase VI and alpha-amylase. Twenty-nine proteins (all porcine) were predominantly secreted by the mandibular and sublingual glands, for example salivary lipocalin and submaxillary apomucin protein.
Project description:Organoids have huge therapeutic potential and are increasingly opening up new avenues within regenerative medicine. However, the clinical application is greatly limited by the lack of effective GMP-complaint systems for organoid expansion in culture. Here, we envisage that the use of extracellular matrix hydrogels derived from decellularized tissues (DT) can provide the instructive native environment necessary to direct cell growth. These gels possess the biochemical signature of tissue-specific ECM, and have the potential for clinical translation due to their natural, non-carcinogenic origin. We show that gels developed from decellularized porcine small intestinal mucosa have similar physiological range and mechanical properties to commercially available gels. ECM-derived gels enable formation and growth of endoderm-derived organoids and, moreover, supports in vivo organoid growth. The development of these ECM-derived hydrogels opens up the potential for human organoids to be used clinically.
Project description:High-resolution shotgun proteomic analysis on two porcine collagen-based biomaterials designed for guided bone regeneration and soft tissue augmentation was perfomed. A porcine-derived collagen membrane and a collagen matrix derived from peritoneum and/or skin were digested, separated by nano-reverse-phase high-performance liquid chromatography, and peptides subjected to mass spectrometric detection and analysis
Project description:Organoids have extensive therapeutic potential and are increasingly opening up new avenues within regenerative medicine. However, the clinical application is greatly limited by the lack of effective GMP-compliant systems for organoid expansion in culture. Here, we envisage that the use of extracellular matrix hydrogels derived from decellularized tissues (DT) can provide an environment capable of directing cell growth. These gels possess the biochemical signature of tissue-specific ECM, and have the potential for clinical translation. Gels from decellularized porcine small intestinal mucosa/submucosa enable formation and growth of endoderm-derived human organoids, such as gastric, liver, pancreas, and small intestine. ECM gels could be used for direct organoids derivation from human biopsies, multiple passages with transcriptomic signature comparable to gold standard culture system, and in vivo organoid delivery. The development of these ECM-derived hydrogels opens up the potential for human organoids to be used clinically.