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Tissue Eng Part C Methods. 2010 Dec;16(6):1621-8. doi: 10.1089/ten.TEC.2010.0146. Epub 2010 Jun 7.

Long-term spatially defined coculture within three-dimensional photopatterned hydrogels.

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Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, Georgia.


Spatially controlled coculture in three-dimensional environments that appropriately mimic in vivo tissue architecture is a highly desirable goal in basic scientific studies of stem cell physiological processes (e.g., proliferation, matrix production, and tissue repair) and in enhancing the development of novel stem-cell-based clinical therapies for a variety of ailments. This study describes a novel fabrication system for photopatterning and assembling cell-laden oligo(polyethylene glycol)-fumarate:poly(ethylene glycol)-diacrylate hydrogels with high spatial fidelity and thickness using a controlled, inert nitrogen environment without the need for expensive precision equipment. Cross-linking was performed using Irgacure-2959 photoinitiator and 365-nm light (∼7 mW/cm²) to form gels ranging from 0.9 to 3 mm in width. Employing a nitrogen environment increased gel thickness up to 240%, generating gels > 1 mm thick before swelling. This technique was further applied for spatially controlled patterning of primary tendon/ligament fibroblasts and marrow stromal cells in a single 1.5-mm-thick laminated hydrogel construct. Cells encapsulated using this technique maintained viability over 14 days in culture. This system potentially enables better understanding of paracrine effects on a range of stem cell functions and therefore may be useful as an in vitro model system for a wide array of regenerative medicine applications.

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