Microfabricated poly(ethylene glycol) templates enable rapid screening of triculture conditions for cardiac tissue engineering
Microfabricated poly(ethylene glycol) templates enable rapid screening of triculture conditions for cardiac tissue engineering
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DOI：
10.1002/jbm.a.32014
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发表时间：
2009-06-01
影响因子：
4.9
通讯作者：
Radisic, Milica
中科院分区：
文献类型：
--
作者：
Iyer, Rohin K.;Chiu, Loraine L. Y.;Radisic, Milica
本研究的目的是设计一个简单的系统用于培养微型心脏类器官，并研究细胞组成对类器官功能的影响。我们假设，与同时接种三种细胞类型或单独培养富集的CM相比，在成纤维细胞（FB）和内皮细胞（EC）的预制网络上培养心肌细胞（CM）将增强类器官的结构和功能特性。通过光聚合聚（乙二醇）二丙烯酸酯来创建用于细胞接种的微通道。在预培养组中，通道用NIH 3 T3 FB和D4 T EC的混合物接种，然后在FB/EC预培养2天后加入新生大鼠CM。对照微通道与FB/EC/CM（同时三育）或单独与富集CM（富集CM）同时接种。预培养产生圆柱形、收缩性和致密的心脏类器官，其中含有表达连接蛋白43和心肌肌钙蛋白I的细长CM。相比之下，同时triculture导致非收缩类器官与集群的CM增长单独从细长的FB和EC。在同时三代培养组中不存在连接蛋白-43的染色。当固定或冷冻FB/EC被用作CM的预培养底物时，获得了非收缩性类器官;而在单细胞类型（FB或EC）上预培养导致收缩性类器官，但与用FB/EC预培养相比具有较差的性质。这些结果强调了活细胞的重要性，两种非肌细胞细胞类型的存在，以及培养功能性多细胞型心脏类器官的顺序接种方法。0 2008 Wiley Periodicals，Inc. J Biomed Mater Res 89A：616-631，2009
The purpose of this study was to design a simple system for cultivation of micro-scale cardiac organoids and investigate the effects of cellular composition on the organoid function. We hypothesized that cultivation of cardiomyocytes (CM) on preformed networks of fibroblasts (FB) and endothelial cells (EC) would enhance the structural and functional properties of the organoids, compared to simultaneously seeding the three cell types or cultivating enriched CM alone. Microchannels for cell seeding were created by photopolymerization of poly(ethylene glycol) diacrylate. In the preculture group the channels were seeded with a mixture of NIH 3T3 FB and D4T EC, following by addition of neonatal rat CM after 2 days of FB/EC preculture. The control microchannels were seeded simultaneously with FB/EC/CM (simultaneous triculture) or with enriched CM alone (enriched CM). Precullure resulted in cylindrical, contractile, and compact cardiac organoids that contained elongated CM expressing connexin-43 and cardiac troponin I. In contrast, simultaneous triculture resulted in noncontractile organoids with clusters of CM growing separately from elongated FBs and ECs. The staining for Connexin-43 was absent in the simultaneous triculture group. When fixed or frozen FB/EC were utilized as a preculture substrate for CM, noncontractile organoids were obtained; while preculture on a single cell type (either FB or EC) resulted in contractile organoids but with inferior properties compared to preculture with both FB/EC. These results emphasize the importance of living cells, presence of both nonmyocyte cell types as well as sequential seeding approach for cultivation of functional multicell type cardiac organoids. 0 2008 Wiley Periodicals, Inc. J Biomed Mater Res 89A: 616-631, 2009