Use of adipose-derived stem cells to fabricate scaffoldless tissue-engineered neural conduits in vitro
| Autor: | Lisa M. Larkin, Ellen M. Arruda, Aaron M. Adams |
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| Rok vydání: | 2012 |
| Předmět: |
Indoles
Neurogenesis Cellular differentiation Population Nerve guidance conduit Biology Fibroblast growth factor Collagen Type I Article Tissue engineering Neurosphere Adipocytes Animals education Cell Proliferation Analysis of Variance education.field_of_study Epidermal Growth Factor Tissue Engineering Stem Cells General Neuroscience S100 Proteins Cell Differentiation Fibroblasts Coculture Techniques Rats Inbred F344 Rats Cell biology Peripheral nerve injury Immunology Female Fibroblast Growth Factor 2 Stem cell |
| Zdroj: | Neuroscience. 201:349-356 |
| ISSN: | 0306-4522 |
| DOI: | 10.1016/j.neuroscience.2011.11.004 |
| Popis: | Peripheral nerve injuries resulting from trauma or disease often necessitate surgical intervention. Although the gold standard for such repairs uses nerve autografts, alternatives that do not require invasive harvesting of autologous nerve tissues are currently being designed and evaluated. We previously established the use of scaffoldless engineered neural conduits (ENCs) fabricated from primary cells as one such alternative in sciatic nerve repair in rats [Baltich et al. (2010) In Vitro Cell Dev Biol Anim 46(5):438–444]. 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 (3-D) scaffoldless tissue-ENC. Addition of ascorbic acid-2-phosphate and fibroblast growth factor (FGF)-2 to the medium induced and differentiated ASCs to a fibroblast lineage in more than 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 expression of S100. ASC-derived fibroblast monolayers and neurospheres were co-cultured to fabricate a 3-D scaffoldless tissue-ENC. 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. |
| Databáze: | OpenAIRE |
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