Institute of Experimental Gene Therapy and Cancer Research, Rostock University Medical Center, 18057, Rostock, Germany.
Department Life, Light and Matter, University of Rostock, 18059, Rostock, Germany.
Cell Mol Life Sci. 2022 Apr 9;79(5):229. doi: 10.1007/s00018-022-04264-8.
Skeletal muscle tissue engineering aims at generating biological substitutes that restore, maintain or improve normal muscle function; however, the quality of cells produced by current protocols remains insufficient. Here, we developed a multifactor-based protocol that combines adenovector (AdV)-mediated MYOD expression, small molecule inhibitor and growth factor treatment, and electrical pulse stimulation (EPS) to efficiently reprogram different types of human-derived multipotent stem cells into physiologically functional skeletal muscle cells (SMCs). The protocol was complemented through a novel in silico workflow that allows for in-depth estimation and potentially optimization of the quality of generated muscle tissue, based on the transcriptomes of transdifferentiated cells. We additionally patch-clamped phenotypic SMCs to associate their bioelectrical characteristics with their transcriptome reprogramming. Overall, we set up a comprehensive and dynamic approach at the nexus of viral vector-based technology, bioinformatics, and electrophysiology that facilitates production of high-quality skeletal muscle cells and can guide iterative cycles to improve myo-differentiation protocols.
骨骼肌组织工程旨在生成生物替代品，以恢复、维持或改善正常的肌肉功能；然而，目前的方案所产生的细胞质量仍然不足。在这里，我们开发了一种基于多种因素的方案，该方案结合了腺病毒（AdV）介导的 MYOD 表达、小分子抑制剂和生长因子处理以及电脉冲刺激（EPS），可有效地将不同类型的人源多能干细胞重编程为具有生理功能的骨骼肌细胞（SMCs）。该方案通过一种新的计算工作流程得到了补充，该流程允许根据转分化细胞的转录组，深入估计和潜在优化生成的肌肉组织的质量。我们还对表型 SMCs 进行了膜片钳钳制，以将它们的生物电特性与其转录组重编程联系起来。总的来说，我们在基于病毒载体的技术、生物信息学和电生理学的交汇点建立了一种全面而动态的方法，该方法有助于产生高质量的骨骼肌细胞，并可以指导迭代循环，以改进肌分化方案。
