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. Author manuscript; available in PMC: 2019 Jun 1.
Published in final edited form as: J Biomed Mater Res A. 2018 Feb 19;106(6):1543–1551. doi: 10.1002/jbm.a.36351

Figure 1. Engineering structurally organized single and multi-layered skeletal muscle tissue.

Figure 1

Using a thermoresponsive nanofabricated substrata (TNFS) and the gel casting method for engineering and transferring anisotropic cell and ECM monolayers, cells are cultured on TNFS for consistent orientation of cellular alignment (red double pointed arrow indicates substrate orientation). Cells deposit anisotropic ECM (green fibers) and form confluent cell sheets within 48 hours. Following casting in gelatin cell sheets are subjected to reduced temperature to trigger thermoresponse of polymer coated substrate and removed from culture dish following gelatin solidification (black arrow indicates direction of casted cell layer). Coherent cell sheets are stacked on other cell sheets with varying orientations and deposited on a new culture surface for experimental conditions, tissue bilayers with anisotropic cell and ECM orientation were cultured in growth or differentiation conditions for up to seven days maintaining anisotropy over the duration of experimental period.