A strategy for fabrication of a three-dimensional tissue construct containing uniformly distributed embryoid body-derived cells as a cardiac patch
A strategy for fabrication of a three-dimensional tissue construct containing uniformly distributed embryoid body-derived cells as a cardiac patch
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DOI：
10.1016/j.biomaterials.2010.04.067
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发表时间：
2010-08-01
期刊：
影响因子：
14
通讯作者：
Sung, Hsing-Wen
中科院分区：
文献类型：
--
作者：
Huang, Chieh-Cheng;Liao, Chen-Kang;Sung, Hsing-Wen
在体外培养包含多层接种细胞的三维（3D）支架对于在体内构建厚且有活力的心脏组织至关重要。胚胎干细胞（ESCs）已被用作心脏修复的替代细胞来源；然而，解离的ESCs在支架中存活率低，且不会形成类胚体（EB）结构。在本研究中，开发了一种旨在在多孔3D组织支架中均匀培养EB衍生细胞（EBDCs）的策略。该策略采用了在热响应水凝胶系统中形成球对称EB的技术、在涂有胶原蛋白的类似水凝胶系统中制备EBDCs细胞片以及制作切片多孔组织支架的技术。将制备好的EBs收集起来并均匀接种在细胞片培养系统中。培养8天后，获得了具有细胞搏动的连续EBDCs细胞片；我们的定量聚合酶链反应（qPCR）和流式细胞术分析表明，细胞片培养系统上涂覆的胶原蛋白可显著增加心脏谱系细胞的数量。然后将制备的EBDC细胞片夹入切片多孔组织支架中。再次培养后，接种的EBDCs在整个支架中均匀重新分布，机械强度显著提高。在支架中生长的一些细胞表达了心脏特异性肌球蛋白重链和α-辅肌动蛋白，而连接蛋白43在细胞边界处清晰表达。需要进一步开展诸如采用纯化技术富集心肌细胞数量等研究，以进一步改进所开发的组织构建体作为生物工程心脏补片。（C）2010爱思唯尔有限公司。保留所有权利。
Growing three-dimensional (3D) scaffolds that contain more than a few layers of seeded cells in vitro is crucial for the creation of thick and viable cardiac tissues in vivo. Embryonic stem cells (ESCs) have been used as an alternative cell source for cardiac repair; however, dissociated ESCs show poor viability in the scaffold and do not form the embryoid body (EB)-like structures. In this study, a strategy intended for cultivating EB-derived cells (EBDCs) uniformly in a porous 3D tissue scaffold was developed. This strategy employed techniques of formation of spherically symmetric EBs in a thermo-responsive hydrogel system, production of cell sheets of EBDCs in a similar hydrogel system coated with collagen and fabrication of sliced porous tissue scaffolds. The prepared EBs were collected and plated evenly in the cell-sheet culture system. After 8 days in culture, a continuous sheet of EBDCs with cell beating was obtained; our qPCR and flow cytometric analyses showed that the collagen-coated on the cell-sheet culture system can significantly enhance the population of cardiac-lineage cells. The produced EBDC sheets were then sandwiched into the sliced porous tissue scaffold. After reculture, the seeded EBDCs were redistributed uniformly throughout the scaffold, with a significant increase in mechanical strength. Cardiac-specific myosin heavy chain and alpha-actinin were expressed for some cells grown in the scaffold, while connexin 43 was clearly expressed at the cell borders. Additional studies such as employing purification techniques to enrich the population of cardiomyocytes are needed to further improve the developed tissue constructs as a bioengineered cardiac patch. (C) 2010 Elsevier Ltd. All rights reserved.