Novel integrated workflow allows production and in-depth quality assessment of multifactorial reprogrammed skeletal muscle cells from human stem cells.

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Faustino D., Brinkmeier H., Logotheti S., Jonitz-Heincke A., Yilmaz H., Takan I., ...Daha Fazla

Cellular and molecular life sciences : CMLS, cilt.79, sa.5, ss.229, 2022 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 79 Sayı: 5
  • Basım Tarihi: 2022
  • Doi Numarası: 10.1007/s00018-022-04264-8
  • Dergi Adı: Cellular and molecular life sciences : CMLS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, Academic Search Premier, Agricultural & Environmental Science Database, CAB Abstracts, Chemical Abstracts Core, Chimica, EMBASE, MEDLINE, Veterinary Science Database
  • Sayfa Sayıları: ss.229
  • Anahtar Kelimeler: Multipotent stem cells, Myogenic differentiation, Adenovector-mediated MYOD expression, Small molecules, Electrical pulse stimulation, Skeletal muscle transcriptomics, Myo-informatics
  • Dokuz Eylül Üniversitesi Adresli: Evet

Özet

Skeletal muscle tissue engineering aims at generating biological substitutes that restore, maintain or improve normal muscle function; however, the quality of cells produced by current protocols remains insufficient. Here, we developed a multifactor-based protocol that combines adenovector (AdV)-mediated MYOD expression, small molecule inhibitor and growth factor treatment, and electrical pulse stimulation (EPS) to efficiently reprogram different types of human-derived multipotent stem cells into physiologically functional skeletal muscle cells (SMCs). The protocol was complemented through a novel in silico workflow that allows for in-depth estimation and potentially optimization of the quality of generated muscle tissue, based on the transcriptomes of transdifferentiated cells. We additionally patch-clamped phenotypic SMCs to associate their bioelectrical characteristics with their transcriptome reprogramming. Overall, we set up a comprehensive and dynamic approach at the nexus of viral vector-based technology, bioinformatics, and electrophysiology that facilitates production of high-quality skeletal muscle cells and can guide iterative cycles to improve myo-differentiation protocols.