Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Trapp, E.
Cultivation of human Schwann cells and subsequent colonisation of the Polymer OPTIMAIX 2D
Humanmedizin; [ Diplomarbeit ] Medical University of Graz; 2014. pp. 99
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Spendel Stephan
- Altmetrics:
- Abstract:
- Background: Peripheral nerves are frequently severely injured as a result of trauma. This causes deficiency in sensory and motor qualities within the affected innervation area. First line therapy in treating peripheral nerve injury accompanied by nervous tissue defect is the autologous nerve graft. This method however is fundamentally flawed for two major reasons. The first being that harvesting nerves results in innervations deficiency at the donor site, and secondly, there is limited availability of appropriate grafting nerves that can be utilised for reconstruction. A tissue engineered nerve construct could represent a solution for these problems. In animal models it was shown that scaffolds containing Schwann cells have highly positive effects on regenerating nerves. To harness these results for treating peripheral nerve injury in humans it is necessary to harvest and culture Schwann cells from human peripheral nerves and finally apply them to a scaffold. Methods: Within this pilot study human Schwann cells were obtained by explant-reexplant technique and cultured from human peripheral nerves. For harvesting human Schwann cells, peripheral nerves were explanted from amputates of patients at the Department of Plastic, Aesthetic and Reconstructive Surgery at the Medical University of Graz. The isolated nerves were dissected from the epineurium and either cut into 1 mm sized pieces or the fascicles itself were pulled out of the nervous explant and cultured under standard conditions. After forming a confluent monolayer of cells, the tissue and respectively the cells of the monolayer were harvested, applied to OPTIMAIX 2D and cultured for another four weeks. Afterwards each polymer was frozen to obtain 7 µm slices at the cryomicrotome. The sections were stained with H&E for detecting cells on the polymer OPTIMAIX 2D. Additionally Schwann cell specific S100 staining was performed to ensure that the H&E positive cells were Schwann cells. Furthermore MIB staining was conducted on all H&E and S100 positive sections, to depict mitosis for indicating culture growth on the polymer OPTIMAIX 2D. Results: In H&E stained sections a sprouting in all nervous tissue colonised polymers was observed, while the polymers colonised with cells harvested from the monolayer did not show any H&E stained cell at all. S100 staining of the nervous tissue colonised polymers showed a sprouting of Schwann cells in all sections. Furthermore, MIB staining was performed for detecting cell mitosis within the S100 positive cells, which depicted proliferating Schwann cells within the sprouting, indicating culture growth on the polymer OPTIMAIX 2D.