Neurogenic differentiation of murine and human adipose-derived stromal cells
Section snippets
Materials and methods
Cell harvest and culture. For isolation of murine ADAS (muADAS) cells, we used BalbC mice (Charles River Laboratories). Animals were housed under standard conditions and the IACUC of Duke University approved all animal procedures. After sacrifice of the mice, we harvested subcutaneous adipose tissue and isolated ADAS cells using a modification of published methods [25]. Briefly, we mechanically dissociated the adipose tissue, performed a digestion with collagenase type I (Sigma), and
ADAS cell characterization
Within 2–3 passages after the initial plating of the primary culture, muADSCs cells appeared as a monolayer of large, flat cells (25– in diameter). As the cells approached confluence, they assumed a more spindle-shaped, fibroblastic morphology. Flow cytometry analysis of muADAS cells at passages 3–5 demonstrated that the cells were negative for CD11b and CD45 (Fig. 1A), cell surface markers associated with hematopoietic cells. In contrast, the majority of muADAS cells expressed CD29 (Fig. 1B
Discussion
Neural tissue has limited potential for intrinsic repair following injury and the identification of cells capable of neuronal differentiation has generated intense interest [1], [2]. Our results demonstrate that murine and human adipose-derived stem cells cultured in the presence of specific soluble mediators possess the ability to undergo morphologic and phenotypic changes consistent with neuronal differentiation. These findings, together with previous studies, indicate that stem and
Acknowledgements
This work was supported by funds provided by the Owen H. Wagensteen, MD Faculty Research Fellowship of the American College of Surgeons, as well as by funds provided by Artecel Sciences, Inc. Dr. Rice serves as a consultant to Artecel Sciences, Inc. Note that portions of this research were presented at the 33rd Annual Meeting of the American Pediatric Surgical Association, May, 2002. The authors thank Alex Freemerman and Ashok Shetty for expert advice during the course of this work.
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