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

Artur Bryja,Hanna Piotrowska-Kempisty,Ana Angelova Volponi,Grzegorz Latosiński,Marta Dyszkiewicz-Konwińska,Julia Spaczyńska,Jamil A Shibli,Maurycy Jankowski,Paul Mozdziak,Bartosz Kempisty,Rut Bryl,Krzysztof Krawiec

doi:10.3390/ani11010015

Abstract

Simple SummaryDomestic 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.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.

Highlights

Domestic pigs express high phylogenetic similarity to humans and are considered as compatible model in biomedical research [1,2]
The cell suspension was filtered through mesh to remove non-dissociated tissue fragments, and isolated cells were centrifugated at 300× g for 8 min
The analysis showed that differently expressed genes can be classified to 56 gene ontology groups

Summary

Introduction

Domestic pigs express high phylogenetic similarity to humans and are considered as compatible model in biomedical research [1,2]. Advanced genetic engineering opened the possibility of genetically modified pigs’ application as tissue and/or organ donors in human xenotransplantation [3,4]. They were used as a host organism for the formation of the heart and vascular architecture, to produce graft-ready artificial vessels [5,6,7]. Both as a model for biomedical research and an animal donor for tissue biopsies [10,11,12]. Our research focuses on the porcine oral mucosa, consisting of two distinct layers (epithelium and fibrous connective tissue layer), with remarkable capacity for repair [13,14,15]

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