1. Assessment of different strategies for scalable production and proliferation of human myoblasts
- Author
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Tan J, Chunwei Li, Bin Tean Teh, Min-Wen Jason Chua, Suet Lee Shirley Ding, Ng Shyh-Chang, Suet‐Mei Crystal Low, Ege Deniz Yildirim, Zongmin Jiang, Kang Yu, and Yan-Jiang Benjamin Chua
- Subjects
Genetic Markers ,0301 basic medicine ,Satellite Cells, Skeletal Muscle ,Cellular differentiation ,Human Embryonic Stem Cells ,musculoskeletal system ,Biology ,Cell fate determination ,adult stem cells ,immortalization ,Muscle Development ,MyoD ,Cell Line ,Myoblasts ,03 medical and health sciences ,0302 clinical medicine ,Cyclin D1 ,Humans ,Regeneration ,Myocyte ,Telomerase reverse transcriptase ,Telomerase ,Cells, Cultured ,Cell Proliferation ,MyoD Protein ,cell fate ,Cyclin-Dependent Kinase 4 ,Cell Differentiation ,Original Articles ,Cell Biology ,General Medicine ,embryonic stem cells ,Cell Transformation, Viral ,Embryonic stem cell ,Cell biology ,cellular differentiation ,030104 developmental biology ,030220 oncology & carcinogenesis ,Original Article ,tissues ,Adult stem cell - Abstract
Objectives 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. Methods We compared the immortalization of primary myoblasts with hTERT, cyclin D1 and CDK4R24C, two chemically defined methods for deriving myoblasts from pluripotent human embryonic stem cells (hESCs), and introduction of viral MyoD into hESC‐myoblasts. Results 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. Conclusions 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.
- Published
- 2019
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