Muscle-macrophage constructs for skeletal muscle repair

仿生骨骼肌组织的成功工程可以创造精确的 肌肉生理学和疾病的模型，并帮助治疗各种肌肉疾病。这个项目 是基于我们最近开发的方法，利用成年大鼠肌源性细胞的3D工程 骨骼肌组织具有与天然肌肉相当的结构和功能特性。 具体来说，我们已经建立了成年大鼠肌源性细胞体外扩增的条件 并成功地利用它们来设计具有收缩能力的骨骼肌组织， 比以前报道的高100倍。重要的是，在一系列全面的初步研究中， 首次显示了体外成体来源的工程化肌肉的自我再生能力， 通过骨骼肌祖细胞的3-D共培养， 来源于骨髓的非极化巨噬细胞。我们建议在这些令人兴奋的结果的基础上 并系统地探索利用三维肌肉-巨噬细胞共培养系统， 具有快速血管和神经元整合能力的收缩和再生肌肉组织 并在体内成功修复骨骼肌损伤。我们将研究：（1）细胞和分子 体外巨噬细胞介导的组织工程化肌肉自我修复的机制;（2）联合应用 巨噬细胞、血管细胞和生物物理线索对体外形成的前体细胞的能力的影响， 血管化的工程化肌肉在体内经历快速血液灌注和功能成熟， 以及（3）巨噬细胞补充和突触发生刺激在体外对能力的作用 肌肉巨噬细胞植入物的功能整合和修复受损的骨骼肌， vivo.成功完成建议的研究，将为日后的申请奠定基础 组织工程方法学应用于人体肌肉修复。此外，我们的新战略，利用 免疫系统细胞作为组织工程植入物中的促再生佐剂可能会发现广泛的 在再生医学领域的应用。

Successful engineering of biomimetic skeletal muscle tissues could allow creation of accurate models of muscle physiology and disease and aid treatment of various muscle disorders. This project is based on our recently developed methods to utilize adult rat myogenic cells for engineering of 3D skeletal muscle tissues with structural and functional properties comparable to those of native muscle. Specifically, we have established conditions for robust in vitro expansion of adult rat myogenic cells and have successfully utilized them to engineer skeletal muscle tissues with contractile capacity 10- 100 fold higher than previously reported. Importantly, in a comprehensive set of preliminary studies we for the first time show that self-regenerative capacity of adult-derived engineered muscle in vitro and survival in vivo can be significantly enhanced by a 3-D co-culture of skeletal muscle progenitors with non-polarized macrophages derived from bone marrow. We propose to build on these exciting results and systematically explore the use of 3D muscle-macrophage co-culture system to create highly contractile and regenerative muscle tissues with the capacity for rapid vascular and neuronal integration and successful repair of skeletal muscle injury in vivo. We will study: (1) the cellular and molecular mechanisms of macrophage mediated self-repair of tissue-engineered muscle in vitro, (2) the combined effects of macrophages, vascular cells, and biophysical cues on the ability of in vitro formed pre- vascularized engineered muscle to undergo rapid blood perfusion and functional maturation in vivo, and (3) the roles of macrophage supplementation and synaptogenic stimulation in vitro upon the ability of muscle-macrophage implants to functionally integrate with and repair damaged skeletal muscle in vivo. Successful completion of the proposed studies will establish foundation for the future applications of tissue engineering methodologies to human muscle repair. Furthermore, our novel strategy to utilize immune system cells as pro-regenerative adjuvants inside tissue-engineered implants may find broad applications in the field of regenerative medicine.