Behaviour of isolated Olfactory Stem Cells within Cerebrospinal Fluid: a Prerequisite for Cell Therapy after Spinal Cord Injury

Objectives A promising treatment to promote the neurological function after spinal cord injury (SCI) could be an autologous cell therapy. Especially the olfactory mucosa (OM) may be a promising cell source because of the presence of olfactory stem cells (OSC). Cells which are planted into the spinal cord injury will come in contact with cerebrospinal fluid (CSF) of the central nervous system (CNS). Among others, the CSF protects the brain and spinal cord from traumata, regulates the chemical environment of the CNS and mediates the intracerebral transport of molecules involved in key processes. To provide maximum safety to the patients it is important to analyse the influence of CSF on the transplanted cells. Therefore, the viability, proliferation, migration and differentiation potential were analyzed by simulating a SC environment by co-culturing OSC with CSF.

Methods Human OM biopsies were collected during corrections of the nasal septum. Olfactory stem cells were isolated using the neurosphere assay and cells were characterised by FACS and CLSM. CSF was collected from healthy individuals who underwent lumbar puncture for diagnostic purpose. The viability and proliferation in contact to CSF was analyzed by AlamarBlue and Calcein-AM/PI staining after different time points. In addition, cell proliferation and migration was analyzed by CytoSmart Life Imaging and scratch assay. Possible spontaneous osteogenic differentiation was determined by Alizarin Red S staining. The neuronal differentiation potential of OSC was tested with and without CSF by using the ß III-tubulin marker.

Results and Conclusion Our study demonstrated that OSC can be purified by neurosphere assay and express stem cell markers such as CD90 and CD105. In contact to CSF cells are viable and proliferate even in high concentrations of CSF over a period of 21 days. No decrease in the cell proliferation or migration rate could be observed in the presence of CSF. When cultivated in higher concentrations of CSF (80%), OSC change their morphology into a more elongated shape. In addition, their differentiation potential into neuron-like cells expressing ßIII-tubulin was enhanced in the presence of CSF (Fig.1). Spontaneous osteogenic differentiation of OSC in CSF did not occur.

The utilisation of OSC may be a prospective therapy for the treatment of spinal cord injury.

Stichwörter Olfactory mucosa, olfactory stem cells, cell therapy, Cerebrospinal Fluid, autologous cell transplantation, spinal cord injury

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Artikel online veröffentlicht:
15. Oktober 2020