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Biopsies of olfactory mucosa on living animals
Olfactory mucosa biopsies were successfully performed on living animals from all genera except mice, with some differences in collection and culture techniques. Biopsied animals under anesthesia could recover within a few hours after surgery without any sign of pain or unwanted side effects. The only undesirable effect observed in all studied genera was nasal bleeding immediately fol-lowing the biopsies, which was rapidly suppressed by applying a sterile gauze in the nose or on the bone window.
Isolation and amplification of cells with fibroblastic morphology from olfactory mucosa biopsies
One to two weeks after biopsy, adherent cells with heterogeneous morphologies from each genus began to grow out of the explants and invaded the culture dish. After 2 passages, cultures became more homogenous and cells exhibited a fibroblastic morphology . Cells from mouse showed morphological changes with increasing passages or dilution associated with a difficulty in amplifying them. A vulnerability to dilution was also observed for all genera during the first steps of amplification, with a decrease in cell proliferation if split ratio exceeded 1:2 .
OE-MSCs from different mammalian genera display features of stemness
After amplification, two stemness and immaturity features that were previously described in human OE-MSCs were assessed: the ability to give rise to spheres and the expression of nestin. When grown under specific appropriate culture conditions, OE-MSCs from all gen-era could generate spheres.For each genus, the entire population of OE-MSCs expressed the nestin protein, a prominent marker of im-maturity, with similar intensities of expression across all genera. Finally, OE-MSCs were successfully transfected to ex-press GFP, a prerequisite for transplantation studies involving cell tracking.
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