A promising strategy to treat sensorineural hearing loss could be an autologous cell therapy. Due to the lifelong neurogenesis the olfactory mucosa (OM) might be an attractive cell source. The neuro-regenerative properties of olfactory multipotent stem cells (OSC) and olfactory ensheathing cells have been successfully demonstrated in animal models. Up to now, there are no precise and standard methods for cell isolation and characterization. In this study we compared different methods to isolate and characterize OSC from human OM. The viability of these cells was analyzed in an autologous plasma clot matrix as a possible carrier matrix for cell transplantation.
Human OM biopsies were collected during corrections of the nasal septum. To isolate OSC different techniques were used such as MACS-Microbeads or differential adhesion method. For characterization surface markers such as CD90, CD105, p75NTR and nestin were analyzed by FACS and CLSM. The neuronal differentiation potential (ß-tubulin expression) was analyzed by CLSM. Viability within the plasma clot was analyzed by calcein-AM/PI staining.
We demonstrated that OSC isolated from OM are closely related to bone marrow human mesenchymal stem cells (BM-hMSC). OSC proliferate faster than BM-hMSC and express typical BM-hMSC surface markers such as CD90 and show a lack of expression of CD45. Expanded cells are able to form neurospheres and differentiate into neuron-like cells expressing ß-tubulin. In addition, these cells stay viable within a 3D-plasma clot and are able to proliferate within this matrix.
The utilisation of OSC may be a prospective therapy for the treatment of SNHL and the plasma clot might be a favourable matrix for cell transplantation strategies.
