New cell therapy applications for the treatment of traumatic spinal cord injuries represent a new therapeutic approach. These include cells of the olfactory mucosa such as the "ensheathing" cells (OEC). OEC can improve the axonal regeneration, remove cellular debris and have angiogenic properties. In this project, human OEC will be isolated, characterized and functionally analyzed. For possible use for neural regeneration after spinal cord trauma an establishment of methods is especially relevant, which allow a realistic clinical implementation. Therefore, it is essential to find the best method for isolation and enrichment, to perform the cell characterization and cell functionality test as a quality control and to establish appropriate cell-carrier system (autologous plasma clot) for transplantation.
After repeated washing, the olfactory biopsy is subjected to an enzymatic treatment with dispase II to loosen the tissue and to separate the olfactory epithelium and the lamina propria. Subsequently, OEC will be isolated by three different methods to clarify which method leads to an optimal yield of OEC. For cell biological characterization, biochemical and immunological methods will be used:
Cell proliferation rate by BrdU proliferation ELISA. Epitope analysis (for low and high rates of cell division) by FACS and immunohistochemistry: nestin (neural stem cells; glial cells and astrocytes), O4 as well as p75NGF (OEC), GFAP (glial cells), ICAM-1 (stem cells), stem cell and leukocyte lineage markers: CD34, CD45, CD90, CD105 as well as Tuj-1 (immature and mature olfactory receptor neurons). Analysis of neutrotrophic, paracrine factors by proteome Profiler array and ELISA: PDGF, neuropeptide Y and S100, NGF, BDNF, etc.). Phagocytosis property by phagocytosis assay (flow cytometry and fluorescencemicroscopie). Behavior of OEC in a Plasmaclot-Matrix (viability, proliferation, secretion of neurotrophic factors, etc.).
-cross sectoral-Type of hazard:
rehabilitationDescription, key words:
ensheating, spinal cord