FIG. 3.
Construct development and neuronal outgrowth. (A) Top panel: Neuronal progenitor cells were cultured in the absence of smooth muscle and failed to morphologically differentiate through 10 days of culture. (A) Bottom panel: Neuronal progenitor cells cultured in the presence of smooth muscle cells demonstrated elongated morphologies by day 5. (B) By day 8, neuronal progenitor cells were observed to migrate from a sphere and to elongate and form networks (a, a′). The formation on neuronal bundles became evident (b, b′) and an extensive surface network was evident (c, c′). (C) The presence of markers for ENCC (p75NTR), contractile smooth muscle cells (Cal D), neurons (β III tubulin), and glial cells (GFAP) in bioengineered innervated constructs was demonstrated by PCR analysis. Scale bars represent 50 μm in A and 100 μm in B. Cal D, caldesmon; ENCC, neural crest-derived cell; GFAP, glial fibrillary acidic protein.