301 THE VOMERONASAL ORGAN AS A NEW SOURCE OF STEM CELLS FOR CELL THERAPY
M. N. Rodrigues A , G. C. Pignatari A , K. E. B. Grondona B , R. C. Carvalho A C and M. A. Miglino AA University of Sao Paulo, Sao Paulo, Sao Paulo, Brazil;
B Federal University of Paraíba, Areia, Paraiba, Brazil;
C Federal University of Maranhão State, Chapadinha, Maranhão, Brazil
Reproduction, Fertility and Development 25(1) 298-298 https://doi.org/10.1071/RDv25n1Ab301
Published: 4 December 2012
Abstract
The vomeronasal organ (VNO), part of the olfactory system, is located at the base of the nasal cavity at both sides of the nasal septum. The VNO has a tubular shape and possesses receptors to detect pheromones that enable animals to modulate social and reproductive behaviour. Its sensory cells show neurogenesis throughout life, supplied by proliferating stem cells situated at the basal layer that migrate to the surface and generate sensory neurons. The aim of this study was to isolate and characterise stem cells from the VNO in a laboratory animal model species, the New Zealand rabbit. Five males were used for cell isolation and cultivation of stem cells. Three culture media were tested: α-MEM, DMEM-F12, and DMEM-high glucose. As typical for stem cells, isolated cells had a fibroblastic shape, were adhered to the plastic, and achieved good convergence. The cells expanded in all 3 culture media at Day 13. At Day 20, they reached sufficient confluency to promote cell growth and trypsinization. Microscopic analysis revealed that confluence and cell growth were best in DMEM-high glucose medium, confirmed by the proliferation assay (MTT). Cultured stem cells differentiated functionally into adipocytes, osteocytes, and chondrocytes. They did not form tumours when injected into immunocompromised nude mice. In flow cytometry, the cultured cells were negative for CD45 and CD34 and positive for CD105, CD90, Oct3/4, and Stro-1, characterising them as mesenchymal cells. In immunocytochemistry, positive staining was observed for Nanog, vimentin, cytokeratin, Oct3/4, and GFAP. In conclusion, the expression of mesenchymal stem cell markers, the nonexpression of endothelial and haematopoietic markers, as well as the classical patterns of differentiation (adipo-, osteo-, and chondrocytes) and growing processes, indicated that these cells were mesenchymal stem cells. Thus, they may have high therapeutic potential for cell therapy in animals with problems related to reproduction and olfactory function.
