Department of Medicine, Lillehei Heart Institute, University of Minnesota, 4-128 CCRB, 2231 6th Street South East, Minneapolis, MN 55455, USA.
Department of Pediatrics, Lillehei Heart Institute, University of Minnesota, Minneapolis, MN 55455, USA.
Stem Cell Reports. 2017 Jul 11;9(1):12-22. doi: 10.1016/j.stemcr.2017.04.022. Epub 2017 May 18.
Recent reports have documented the differentiation of human pluripotent stem cells toward the skeletal myogenic lineage using transgene- and cell purification-free approaches. Although these protocols generate myocytes, they have not demonstrated scalability, safety, and in vivo engraftment, which are key aspects for their future clinical application. Here we recapitulate one prominent protocol, and show that it gives rise to a heterogeneous cell population containing myocytes and other cell types. Upon transplantation, the majority of human donor cells could not contribute to myofiber formation. As a proof-of-principle, we incorporated the inducible PAX7 lentiviral system into this protocol, which then enabled scalable expansion of a homogeneous population of skeletal myogenic progenitors capable of forming myofibers in vivo. Our findings demonstrate the methods for scalable expansion of PAX7 myogenic progenitors and their purification are critical for practical application to cell replacement treatment of muscle degenerative diseases.
最近的报告记录了使用无转基因和细胞纯化方法将人类多能干细胞向骨骼肌谱系分化。尽管这些方案产生了肌细胞，但它们尚未证明其可扩展性、安全性和体内植入性，这些都是其未来临床应用的关键方面。在这里，我们回顾了一个突出的方案，并表明它产生了一种包含肌细胞和其他细胞类型的异质细胞群体。在移植后，大多数人类供体细胞不能有助于肌纤维的形成。作为一个原理证明，我们将诱导型 PAX7 慢病毒系统纳入该方案，从而能够大规模扩增能够在体内形成肌纤维的同质骨骼肌祖细胞群体。我们的研究结果表明，用于可扩展扩增 PAX7 肌源性祖细胞的方法及其纯化对于将其应用于肌肉退行性疾病的细胞替代治疗至关重要。
