Transcriptomic and Morphological Analysis of Cells Derived from Porcine Buccal Mucosa—Studies on an In Vitro Model

SIMPLE SUMMARY: Domestic pigs express high phylogenetic similarity to humans and are often used as a compatible model in biomedical research. Porcine tissues are used as an accessible biomaterial in human skin transplants and tissue architecture reconstruction. We used transcriptional analysis to investigate the dynamics of complex biological system of the mucosa. Additionally, we performed computer analysis of microscopic images of cultured cells in vitro. Computer analysis of images identified epithelial cells and connective tissue cells in in vitro culture. ABSTRACT: Transcriptional analysis and live-cell imaging are a powerful tool to investigate the dynamics of complex biological systems. In vitro expanded porcine oral mucosal cells, consisting of populations of epithelial and connective lineages, are interesting and complex systems for study via microarray transcriptomic assays to analyze gene expression profile. The transcriptomic analysis included 56 ontological groups with particular focus on 7 gene ontology groups that are related to the processes of differentiation and development. Most analyzed genes were upregulated after 7 days and downregulated after 15 and 30 days of in vitro culture. The performed transcriptomic analysis was then extended to include automated analysis of differential interference contrast microscopy (DIC) images obtained during in vitro culture. The analysis of DIC imaging allowed to identify the different populations of keratinocytes and fibroblasts during seven days of in vitro culture, and it was possible to evaluate the proportion of these two populations of cells. Porcine mucosa may be a suitable model for reference research on human tissues. In addition, it can provide a reference point for research on the use of cells, scaffolds, or tissues derived from transgenic animals for applications in human tissues reconstruction.