Department of Human Genetics, Leiden University Medical Center, 2333 ZA Leiden, the Netherlands.
Department of Clinical Genetics, Erasmus University Medical Center, 3015 GE Rotterdam, the Netherlands; Department of Pediatrics, Erasmus University Medical Center, 3015 GE Rotterdam, the Netherlands; Center for Lysosomal and Metabolic Diseases, Erasmus University Medical Center, 3015 GE Rotterdam, the Netherlands.
Stem Cell Reports. 2023 Oct 10;18(10):1954-1971. doi: 10.1016/j.stemcr.2023.08.014. Epub 2023 Sep 28.
Skeletal muscle research is transitioning toward 3D tissue engineered in vitro models reproducing muscle's native architecture and supporting measurement of functionality. Human induced pluripotent stem cells (hiPSCs) offer high yields of cells for differentiation. It has been difficult to differentiate high-quality, pure 3D muscle tissues from hiPSCs that show contractile properties comparable to primary myoblast-derived tissues. Here, we present a transgene-free method for the generation of purified, expandable myogenic progenitors (MPs) from hiPSCs grown under feeder-free conditions. We defined a protocol with optimal hydrogel and medium conditions that allowed production of highly contractile 3D tissue engineered skeletal muscles with forces similar to primary myoblast-derived tissues. Gene expression and proteomic analysis between hiPSC-derived and primary myoblast-derived 3D tissues revealed a similar expression profile of proteins involved in myogenic differentiation and sarcomere function. The protocol should be generally applicable for the study of personalized human skeletal muscle tissue in health and disease.
骨骼肌研究正朝着 3D 组织工程体外模型转变，这些模型可以再现肌肉的固有结构，并支持对其功能的测量。人类诱导多能干细胞（hiPSCs）可提供大量用于分化的细胞。但是，很难从 hiPSCs 中分化出高质量、纯 3D 的肌肉组织，这些组织表现出的收缩特性可与原代成肌细胞衍生的组织相媲美。在这里，我们提出了一种无转基因方法，可从无饲养层条件下生长的 hiPSCs 中生成纯化、可扩增的成肌祖细胞（MPs）。我们确定了一个最佳的水凝胶和培养基条件的方案，该方案可产生具有类似于原代成肌细胞衍生组织的高收缩性 3D 组织工程骨骼肌。hiPSC 衍生和原代成肌细胞衍生的 3D 组织之间的基因表达和蛋白质组学分析显示，参与成肌分化和肌节功能的蛋白质表达谱相似。该方案应可普遍适用于研究健康和疾病状态下的个性化人类骨骼肌组织。
