NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore City, Singapore.
Stem Cell & Regenerative Biology, Genome Institute of Singapore, Agency for Science Technology and Research, Singapore City, Singapore.
Cell Prolif. 2019 May;52(3):e12602. doi: 10.1111/cpr.12602. Epub 2019 Mar 19.
Myoblast transfer therapy (MTT) is a technique to replace muscle satellite cells with genetically repaired or healthy myoblasts, to treat muscular dystrophies. However, clinical trials with human myoblasts were ineffective, showing almost no benefit with MTT. One important obstacle is the rapid senescence of human myoblasts. The main purpose of our study was to compare the various methods for scalable generation of proliferative human myoblasts.
We compared the immortalization of primary myoblasts with hTERT, cyclin D1 and CDK4 , two chemically defined methods for deriving myoblasts from pluripotent human embryonic stem cells (hESCs), and introduction of viral MyoD into hESC-myoblasts.
Our results show that, while all the strategies above are suboptimal at generating bona fide human myoblasts that can both proliferate and differentiate robustly, chemically defined hESC-monolayer-myoblasts show the most promise in differentiation potential.
Further efforts to optimize the chemically defined differentiation of hESC-monolayer-myoblasts would be the most promising strategy for the scalable generation of human myoblasts, for applications in MTT and high-throughput drug screening.
肌母细胞移植疗法（MTT）是一种用基因修复或健康的肌母细胞替换肌肉卫星细胞的技术，用于治疗肌肉萎缩症。然而，用人肌母细胞进行的临床试验没有效果，MTT 几乎没有带来益处。一个重要的障碍是人类肌母细胞的快速衰老。我们研究的主要目的是比较各种可规模化产生增殖性人类肌母细胞的方法。
我们比较了 hTERT、cyclin D1 和 CDK4 对原代肌母细胞的永生化，以及两种从多能性人类胚胎干细胞（hESC）中获得肌母细胞的化学定义方法，以及将病毒 MyoD 导入 hESC-肌母细胞。
我们的结果表明，虽然上述所有策略在生成既能增殖又能强有力分化的真正人类肌母细胞方面都不理想，但化学定义的 hESC-单层-肌母细胞在分化潜力方面显示出最大的希望。
进一步努力优化化学定义的 hESC-单层-肌母细胞分化将是最有前途的规模化生成人类肌母细胞的策略，可用于 MTT 和高通量药物筛选。
