Yan Wentao, George Sheela, Fotadar Upinder, Tyhovych Natalia, Kamer Angela, Yost Michael J, Price Robert L, Haggart Charles R, Holmes Jeffrey W, Terracio Louis
Department of Basic Sciences, New York University, New York, New York 10010, USA.
Tissue Eng. 2007 Nov;13(11):2781-90. doi: 10.1089/ten.2006.0408.
Loss of skeletal muscle profoundly affects the health and well-being of patients, and there currently is no way to replace lost muscle. We believe that a key step in the development of a prosthesis for reconstruction of dysfunctional muscular tissue is the ability to reconstitute the in vivo-like 3-dimensional (3D) organization of skeletal muscle in vitro with isolated satellite cells. In our present proof of principle studies, we have successfully constructed a multilayered culture of skeletal muscle cells, derived from neonatal satellite cells, that are distributed in a 3D pattern of organization that mimics many of the features of intact tissue. These multilayered cultures are composed of elongated multinucleated myotubes that are MyoD positive. Histological studies indicate that the multiple layers of myotubes can be distinguished. Expression of muscle-specific markers such as myosin heavy chain, dystrophin, integrin alpha-7, alpha-enolase, and beta-enolase was detected using real-time reverse transcriptase polymerase chain reaction at levels near adult values. Physiological measurements of the engineered skeletal muscle showed that they tetanize and display physiologic force length behavior, although developed force per cross-sectional area was below that of native rat skeletal muscle.
骨骼肌的丧失会严重影响患者的健康和福祉，目前尚无办法替代丧失的肌肉。我们认为，开发用于重建功能失调肌肉组织的假体的关键一步是能够在体外利用分离的卫星细胞重建类似体内的骨骼肌三维（3D）组织结构。在我们目前的原理验证研究中，我们成功构建了源自新生卫星细胞的骨骼肌细胞多层培养物，这些细胞以3D组织模式分布，模拟了完整组织的许多特征。这些多层培养物由呈MyoD阳性的细长多核肌管组成。组织学研究表明，可以区分多层肌管。使用实时逆转录聚合酶链反应检测到肌肉特异性标志物如肌球蛋白重链、抗肌萎缩蛋白、整合素α-7、α-烯醇化酶和β-烯醇化酶的表达水平接近成年值。对工程化骨骼肌的生理测量表明，它们会发生强直收缩并表现出生理力长度行为，尽管每横截面积产生的力低于天然大鼠骨骼肌。
