Muscle-macrophage constructs for skeletal muscle repair

成功的仿生骨骼肌组织工程可以创造出准确的 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利用成年大鼠肌源性细胞进行3D工程的方法 结构和功能特性可与天然肌肉相媲美的骨骼肌组织。 具体地说，我们已经为成年大鼠肌源性细胞的体外扩增创造了条件。 并成功地利用它们来设计具有收缩能力的骨骼肌组织10- 比之前报道的高出100倍。重要的是，在一套全面的初步研究中，我们 首次显示成人来源的工程化肌肉的体外和自我再生能力 骨骼肌祖细胞与骨骼肌祖细胞三维共培养可显著提高体内存活率 来自骨髓的非极化巨噬细胞。我们打算在这些令人兴奋的结果的基础上再接再厉 并系统探索利用3D肌肉-巨噬细胞共培养系统创造高度 具有快速血管和神经元整合能力的收缩和再生肌肉组织 并在活体内成功修复骨骼肌损伤。我们将研究：(1)细胞和分子 巨噬细胞介导的组织工程肌肉体外自我修复机制，(2)联合 巨噬细胞、血管细胞和生物物理信号对体外预成纤维细胞能力的影响 血管化的工程化肌肉在体内经历快速血液灌流和功能成熟， (3)巨噬细胞补充和体外突触刺激对能力的影响 肌肉-巨噬细胞植入物与受损骨骼肌功能整合和修复的研究 活着。成功完成拟议的研究将为今后的申请奠定基础 将组织工程方法应用于人体肌肉修复。此外，我们的新战略将利用 在组织工程植入物中，免疫系统细胞作为促再生佐剂可能会发现 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.