In vitro Maintenance of Olfactory Mucosa: with Enriched Olfactory Ensheathing Cells

Abstract

Human Olfactory Mucosa (OM) regulates olfaction through axonal regeneration and myelination mediated by stem cells and Olfactory Ensheathing Cells (OECs) resident in the niche. Purified OECs/olfactory biopsies have been utilized for functional recovery in different Spinal Cord Injury (SCI) models. However, recent reports find this debatable where we propose primary culture of OM, basal cells of olfactory epithelium and olfactory ecto-mesenchymal stem cells. Our defined culture conditions improve the life span of OM with enrichment of OECs providing a strategy for employment for SCI/cochlear damage repair.Briefly, OM post-collection, was non-enzymatically sliced, cultured for 6 weeks and cells characterized morphologically, immuno-cytochemically and western blotting. By day 21, ~70% GFAP and p75NTR stained, spindle shaped astrocyte-like and flattened sheet-like OECs displayed axonal remyelination. By day 30, caspase 3, 8, 9 (gene-product and activity), phospho-p53 negative; GFAP and p75NTR positive dense, overlapping mass of cells was found. This was accompanied with degenerative changes by 6 weeks through GFAP staining. Conversely, trypsination on day 21 resulted in >95% OECs with flattened morphology, GFAP and p75NTR positivity. The human derived OECs were compared with the 2-day SD rat Olfactory Bulb Cells cultured for 2 weeks in F12 media (GFAP and p75NTR positive). Hence, cultured olfactory mucosa displaying axonal regeneration with OECs in culture provides a vehicle for SCI/cochlear damage repair studies.

Effectiveness of bone marrow-derived mononuclear stem cells for neurological recovery in participants with spinal cord injury: A randomized controlled trial

“…[3] We have already conducted a study in support of this for neurogenesis enhancement and axonal re-myelination using olfactory ensheathing cells under defined media. [4] Further, our group had worked on ways to stimulate stem cells to regenerate neurons for functional recovery with promising results.…”

Rat Olfactory Mucosa Mesenchymal Stem/Stromal Cells (OM-MSCs): A Characterization Study

“…In the period between 1 and 2 weeks a er plating of explants, it was possible to observe irradiant spindle-shaped cells in the plate, as detected in other works performed with rat [22] and other species [34,46]. Although some authors prefer a simple method without performing enzymatic digestion of the OM fragments [28,53], the enzymatic treatment before plating the explants did not affect the characteristics of OM-MSCs and was corroborated by other authors [30]. A er isolation of OM-MSCs and their culture, it was possible to observe their features of MSCs [10], namely plastic adhesion and fibroblastic-like shape, as confirmed by other authors [29,54].…”