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Utilization of Adipose-Derived Stem Cells to Fabricate Scaffold-less Tissue Engineered Neural Conduits
Aaron Adams, PhD; Ellen M Arruda; Lisa M. Larkin
University of Michigan, Ann Arbor, MI, USA
Peripheral nerve injuries resulting from trauma or disease often necessitate surgical intervention. While the gold standard for such repairs utilizes nerve autografts, alternatives that do not require the invasive harvesting of autologous nerve tissues are currently being designed and evaluated. We previously established the use of scaffold-less engineered neural conduits fabricated from primary cells as one such alternative in sciatic nerve repair technologies in rats (publication pending). The present study establishes protocols for fabricating neural conduits from adipose-derived stem cells (ASCs) differentiated to either a fibroblast or neural lineage and co-cultured into a three-dimensional scaffold-less tissue engineered neural conduit. Addition of ascorbic acid-2-phosphate with proline and fibroblast growth factor FGF-2 to the medium induced and differentiated ASCs to a fibroblast lineage in over 90% of the cell population, as confirmed by collagen I expression. ASC-differentiated fibroblasts formed monolayers, delaminated and formed 3-D conduits. Neurospheres were formed by culturing ASCs on non-adherent surfaces in serum-free neurobasal medium with the addition of epidermal growth factor (EGF) and FGF-2. The addition of 10 ng EGF and 10 ng FGF-2 produced larger and more numerous neurospheres than treatments of lower EGF and FGF-2 concentrations. Subsequent differentiation to glial-like cells was confirmed by the presence of S100 marker. ASC-derived fibroblast monolayers and neurospheres were co-cultured to fabricate a three-dimensional scaffold-less tissue engineered neural conduit. Their nerve-like structure and incorporation of glial-like cells which would associate with regenerating axons may make these novel, stem cell derived neural conduits an efficacious technology for repairing critical gaps following peripheral nerve injury.
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