Artikel
Adult human schwann cells: analysis of in vivo potential as a cellular basis for biohybrid nerve grafts for peripheral nerve reconstruction
Die Verwendung von adulten humanen Schwann-Zellen als zelluläre Komponente in biohybriden Transplantaten zur Rekonstruktion peripherer Nerven
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Autoren
Veröffentlicht: | 30. Mai 2008 |
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Gliederung
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Objective: Aim of the study was the evaluation of adult human Schwann cells (ahSCs) as a promising candidate for cell-based peripheral nerve reconstruction therapy in vivo. Regeneration parameters were qualitatively and quantitatively assessed together with investigating the fate of ahSCs after implantation in a long peripheral nerve gap and their involvement in re-establishment of functional nerve fibers.
Methods: Naive (physiological) ahSCs isolated from nerve biopsies (male, 15, 23 & 51yrs) were enriched in vitro [1]. 7x105 cells were either labeled with PKH-26-GL (fluorescent cell surface) or left unlabeled prior to re-suspension in Matrigel before transplantation in silicone tubes to bridge 10 mm sciatic nerve gaps. Immunosuppression (Cyclosporine A) was maintained over the complete experiment to avoid graft rejection. 2, 4 and 6 weeks after transplantation, PKH-26-GL labeled ahSCs were analyzed with regard to survival, distribution and localization pattern within regenerated nerve tissue (n=3, each) and functional behavior (myelination of regenerated axons). 3 and 7 weeks after transplantation of unlabeled ahSCs (n=6, each), regenerated nerve tissues were epon embedded and histomorphometrically analyzed for the number of regenerated myelinated axons in comparison to acellular control filled with Matrigel alone (n=5, each).
Results: PKH-26-GL labeled ahSCs survived and were found to be heterogeneously distributed throughout the regenerated tissue cable in a distinct pattern. AhSCs demonstrated close association with the regenerating axons only 6 weeks after transplantation. Number of nerve transplants including gap-bridging tissue cables after 3 and 7 weeks was significantly higher after ahSC transplantation as compared to acellular control. Histomorphometry showed enhanced overall regeneration parameters 7 weeks after transplantation of ahSCs.
Conclusions: The current study demonstrates for the first time that ahSCs grafted into a long gap in a peripheral nerve survive and show gap-wide distribution in a definite pattern. AhSCs seems to be associated with regenerating axons and point towards a role in supporting axonal regeneration.
References
- 1.
- Haastert K, Mauritz C, Chaturvedi S, Grothe C. Human and rat adult Schwann cell cultures: fast and efficient enrichment and highly effective non-viral transfection protocol. Nat Protoc. 2007;2(1):99-104.