Methodology for the formation of functional, cell-based cardiac pressure generation constructs in vitro
Methodology for the formation of functional, cell-based cardiac pressure generation constructs in vitro
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DOI：
10.1007/s11626-008-9098-9
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发表时间：
2008-10-01
影响因子：
2.1
通讯作者：
Brown, David L.
中科院分区：
文献类型：
--
作者：
Birla, Ravi K.;Dow, Douglas E.;Brown, David L.
我们之前已经描述了一个模型，在体外工程三维（3-D）心肌。在目前的研究中，我们扩展了我们的三维心肌模型，以设计一种基于功能细胞的心脏压力生成结构（CPGC）。利用相分离方法以壳聚糖为支架材料制备管状结构。将从大鼠心脏分离的原代心脏细胞铺在浇铸在35 mm组织培养皿中的纤维蛋白凝胶表面上。通过将管状结构锚定到板的中心形成CPGC（N=8），其中原代心脏细胞接种在包裹管状结构的纤维蛋白凝胶中。在有和没有外部电刺激的情况下评价腔内压力测量值，并进行组织学评价。纤维蛋白凝胶由于心脏细胞的牵引力而自发地压缩。在原始细胞铺板后14天，心肌细胞已完全形成围绕管状构建体的单层，从而形成基于细胞的CPGC。肉眼可见CPGC的自发收缩性，并导致0.08 mmHg的腔内压力峰值。在电刺激时，CPGC产生高达0.05 mmHg的抽搐压力。此外，CPGC结构以高达3 Hz的频率进行电起搏。组织学评价显示在管状结构表面周围存在连续的细胞单层。在这项研究中，我们描述了一种新的体外方法，工程功能细胞为基础的CPGC和功能性能的几个生理指标。
We have previously described a model to engineer three-dimensional (3-D) heart muscle in vitro. In the current study, we extend our model of 3-D heart muscle to engineer a functional cell-based cardiac pressure generating construct (CPGC). Tubular constructs were fabricated utilizing a phase separation method with chitosan as the scaffolding material. Primary cardiac cells isolated from rat hearts were plated on the surface of fibrin gels cast in 35 mm tissue culture dishes. CPGCs (N=8) were formed by anchoring the tubular constructs to the center of the plate with primary cardiac cells seeded in fibrin gels wrapped around the tubular constructs. Intraluminal pressure measurements were evaluated with and without external electrical stimulation and histological evaluation performed. The fibrin gel spontaneously compacted due to the traction force of the cardiac cells. By 14 d after original cell plating, the cardiac cells had completely formed a monolayer around the tubular construct resulting in the formation of a cell-based CPGC. The spontaneous contractility of the CPGC was macroscopically visible and resulted in intraluminal pressure spikes of 0.08 mmHg. Upon electrical stimulation, the CPGCs generated twitch pressures of up to 0.05 mmHg. In addition, the CPGC constructs were electrically paced at frequencies of up to 3 Hz. Histological evaluation showed the presence of a continuous cell monolayer around the surface of the tubular construct. In this study, we describe a novel in vitro method to engineer functional cell-based CPGCs and demonstrate several physiological metrics of functional performance.