Differentiation of Rabbit Stem Cells on Gelatin for Tissue Engineering Applications

3D bioprinting is a novel technology allowing for the precise and controlled deposition of cells and extracellular matrix in order to manufacture artificial tissues. Gelatin is a material commonly used for various bioprinting procedures, owing to its suitability for cell attachment, biocompatibility and optimal physical properties for the printing procedure. In this research, we aimed to compare the smooth muscle and endothelial differentiation potential of rabbit adipose (ASC) and buccal mucosa (BMSC) stem cells cultivated on gelatin. These types of cells could be used in various tissue engineering applications, such as the bioprinting of larger blood vessels or urethral tissue. In the case of smooth muscle differentiation, both types of cells (ASC and BMSC) show reduced proliferation rates indicating commitment to differentiation. Both cell types showed similar patterns of smooth muscle differentiation-related gene expression: ASC and BMSC displayed increased intensity of Acta2 and Tagln gene expression 10 days after differentiation induction; ASC also had increased level of Cald1 expression 5 days after differentiation induction. The appearance of alpha smooth muscle actin stress fibers in ASC cells days after differentiation induction was further confirmed by immunocytochemistry. In the case of epithelial differentiation, both cell types showed similar changes in proliferation intensity 10 days after differentiation induction, as well as no clear differences in gene expression patterns. Thus, we conclude that both stem cell types, when cultivated on gelatin, are similarly suitable for epithelial differentiation, however, ASC would be more suitable for smooth muscle differentiation.