Analysis of olfactory stem- and ensheathing-cells as well as mesenchymal stem cells in contact with cerebrospinal fluid: prerequisite for human cell therapeutic approaches after spinal cord trauma

Status:

ongoing

Aims:

A potential strategy for the treatment of spinal cord injuries represents the cell therapy with olfactory ensheathing- or stem cells as well as with mesenchymal stem cells. Such cell therapies could enable the replacement or support of injured glial cells and neurons. The techniques of cell transplantations are different. However, one important common feature is that all cells will be in contact with cerebrospinal fluid (CSF). For this reason, this project clarifies in advance what effect the CSF may have on the transplanted cells in order to identify possible adverse reactions (such as osteogenic differentiation) in addition to therapeutically desired effects.

Optimal isolation and purification methods for olfactory stem- and ensheathing-cells have been identified. Here, the influence of CSF on the cells, such as on the viability, proliferation and also cell functionality should be analysed.

Activities/Methods:

For the planned project, the olfactory cells should be enriched in complex procedures from nasal mucosal biopsies. These techniques are already established. Mesenchymal stem cells are purchased commercially. Cerebrospinal fluid is provided by the cooperation partner of neurosurgery and neurotraumatology (Prof. Martínez-Olivera). The following methods are used:

• Cell viability of human cells in CSF using calcein-AM/DAPI /Pi staining and the CytoSmart Live Imaging System
• Proliferation behavior of human cells in CSF (BrdU proliferation ELISA and flow cytometric analysis of expression of Ki67)
• Cell typing (CD34, CD45, CD90, CD105, Stro-1, GFAP and p75, S100, Nestin and O4) using FACS
• Analysis of cell functionality in CSF (cell migration, phagocytosis, differentiation etc)
• Analysis of the release of neurotrophic factors of cells in CSF (Human Growth Factor Antibody Array and ELISA).

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