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Layer-by-Layer Assembly for Fabrication of Three-Dimensional Layered Tissues

Speaker : Prof. Michiya Matsusaki
Date & Time : 2009. 6. 16 (Tue), 16:00
Place : 신소재공학과 #2425강의실

Layer-by-Layer Assembly for Fabrication of Three-Dimensional Layered Tissues

The construction of artificial three- dimensional (3D) tissues possessing a similar structure and functions as natural tissue is a key challenge for implantable tissues in tissue engineering, and for model tissues in pharmaceutical assay. The cells are surrounded by nanometer-sized extracellular matrix (ECM) fibers and the fibrous meshwork of ECM dynamically controls a microenvironment of cell surface for adhesion, differentiation, morphology, and organization. The control of microenvironment on cell surface like ECM is attractive and challenging for not only regulation of cell properties and functions but also creation of three-dimensional (3D) engineered tissues.

Recently, we demonstrated a simple and hierarchical cell manipulation technique for the construction of hierarchically layered tissues, which possess a precisely controlled layer number and type of cells, by the direct fabrication of a fibronectin-gelatin (FN-G) nanofilm like an ECM on the surface of each cell layer (Figure 1)1. The FN-G nanofilms were prepared on the cell surface by layer-by-layer (LbL) assembly. The LbL technique is an appropriate method to prepare nanometer-sized films on a substrate through alternative immersion into interactive polymer solutions2. Since the FN-G nanofilms with over 6 nm thick acted as a scaffold for the cell adhesion of the second layer, we successfully developed hierarchical cellular multilayers composed of various type of cells, such as human primary fibroblasts, mouse C2C12 myoblasts, rat primary cardiac myocytes, and human primary smooth muscle cells, even 10 layers (Figure 2). In addition, blood vessel model tissues with varied layer numbers composed of human smooth muscle cell (UASMC) and human endothelial cell (HUVEC) were also fabricated by this technique. These hierarchical structures would be useful as a 3D-layered model tissue for tissue engineering and pharmaceutical assay.

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