FIG. 2.

Cellular morphology and protein expression characteristics of BAM-supported tissue engineered muscle repair (TEMR) constructs developed under three distinct culture conditions. TEMR-1SP, TEMR-1SPD, and TEMR-2SPD constructs are depicted in (A), (B), and (C), respectively (400× images). For the generation of the TEMR-2SPD constructs, a second batch of MDCs was added to an underlying layer of MDCs (i.e., TEMR-1SPD constructs). To confirm adherence of the second MDC batch, these cells were loaded with cytoplasmic fluorescent dye and then visualized following preconditioning (D and E). Scale bar=50 μm for all images. The number of nuclei (F) and the number of multinucleated cells (G) were quantitated for each construct type (See Methods, *TEMR-1SP; ^#TEMR-1SPD, p<0.05). Muscle-specific protein expression of TEMR constructs was characterized via Western blot (H). The optical densities of specified proteins were normalized to that of GAPDH for statistical comparisons among groups [(I); *significantly different from TEMR-1SP, p<0.05]. Protein expression of each construct type is summarized (J). Color images available online at www.liebertonline.com/tea