Long-Term Spatially Defined Coculture Within Three-Dimensional Photopatterned Hydrogels
Long-Term Spatially Defined Coculture Within Three-Dimensional Photopatterned Hydrogels
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DOI：
10.1089/ten.tec.2010.0146
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发表时间：
2010-12-01
影响因子：
3
通讯作者：
Temenoff, Johnna S.
中科院分区：
文献类型：
--
作者：
Hammoudi, Taymour M.;Lu, Hang;Temenoff, Johnna S.
在适当模拟体内组织结构的三维环境中进行空间控制的共培养是干细胞生理过程的基础科学研究中非常期望的目标（例如，例如，在一个实施例中，增殖、基质产生和组织修复），以及促进用于多种疾病的基于干细胞的新型临床疗法的开发。本研究描述了一种新的制造系统，用于使用受控的惰性氮气环境，在不需要昂贵的精密设备的情况下，对具有高空间保真度和厚度的载有细胞的寡聚（聚乙二醇）-富马酸酯：聚（乙二醇）-二丙烯酸酯水凝胶进行组装。使用Irgacure-2959光引发剂和365 nm光（类似于7 mW/cm（2））进行交联，形成宽度为0.9至3 mm的凝胶。采用氮气环境使凝胶厚度增加高达240%，在溶胀之前产生>1 mm厚的凝胶。该技术进一步应用于单个1.5 mm厚的层压水凝胶结构中的原代肌腱/韧带成纤维细胞和骨髓基质细胞的空间控制图案化。使用该技术包封的细胞在培养物中保持活力超过14天。该系统可能使更好地理解旁分泌对一系列干细胞功能的影响，因此可能是有用的，作为一个体外模型系统，广泛的再生医学应用。
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 (similar to 7 mW/cm(2)) to form gels ranging from 0.9 to 3mm 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.