Gholobova Dacha, Gerard Melanie, Terrie Lisanne, Desender Linda, Shansky Janet, Vandenburgh Herman, Thorrez Lieven
Tissue Engineering Laboratory, Department of Development and Regeneration, KU Leuven campus Kulak, Kortrijk, Belgium.
Department of Pathology and Laboratory Medicine, Brown University, Providence, RI, USA.
Methods Mol Biol. 2019;1889:169-183. doi: 10.1007/978-1-4939-8897-6_10.
Skeletal muscle tissue engineering aims at creating functional skeletal muscle in vitro. Human muscle organoids can be used for potential applications in regenerative medicine, but also as an in vitro model for myogenesis or myopathology. However, the thickness of constructs is limited due to passive diffusion of nutrients and oxygen. Introduction of a vascular network in vitro may solve this limitation. Here, we describe tissue engineering of in vitro skeletal muscle consisting of human aligned myofibers with interspersed endothelial networks. To create bio-artificial muscle (BAM), human muscle progenitor cells are cocultured with human umbilical vein endothelial cells (HUVECs) in a fibrin hydrogel. The cell-gel mix is cast into silicone molds with end attachment sites and cultured in endothelial growth medium (EGM-2) for 1 week. The passive forces generated in the contracted hydrogel align the myogenic cells parallel to the long axis of the contracted gel such that they fuse into aligned multinucleated myofibers. This results in the formation of a 2 cm long and ~1.5 mm tick human BAM construct with endothelial networks.
骨骼肌组织工程旨在体外构建功能性骨骼肌。人类肌肉类器官可用于再生医学的潜在应用，也可作为肌发生或肌病理学的体外模型。然而，由于营养物质和氧气的被动扩散，构建物的厚度受到限制。在体外引入血管网络可能解决这一限制。在此，我们描述了由人排列的肌纤维和散布的内皮网络组成的体外骨骼肌组织工程。为了创建生物人工肌肉（BAM），将人类肌肉祖细胞与人脐静脉内皮细胞（HUVECs）在纤维蛋白水凝胶中共培养。将细胞 - 凝胶混合物浇铸到带有末端附着位点的硅胶模具中，并在内皮生长培养基（EGM - 2）中培养1周。收缩水凝胶中产生的被动力使成肌细胞平行于收缩凝胶的长轴排列，从而融合成排列的多核肌纤维。这导致形成一个长2厘米、厚约1.5毫米且带有内皮网络的人类BAM构建物。
