Rossi, Rachele;
Torelli, Silvia;
Ala, Pierpaolo;
Weston, William;
Morgan, Jennifer;
Malhotra, Jyoti;
Muntoni, Francesco;
(2023)
MyoD-induced reprogramming of human fibroblasts and urinary stem cells in vitro: protocols and their applications.
Frontiers in Physiology
, 14
, Article 1145047. 10.3389/fphys.2023.1145047.
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Abstract
The conversion of fibroblasts into myogenic cells is a powerful tool to both develop and test therapeutic strategies and to perform in-depth investigations of neuromuscular disorders, avoiding the need for muscle biopsies. We developed an easy, reproducible, and high-efficiency lentivirus-mediated transdifferentiation protocol, that can be used to convert healthy donor fibroblasts and a promising new cellular model, urinary stem cells (USCs), into myoblasts, that can be further differentiated into multinucleated myotubes in vitro. Transcriptome and proteome profiling of specific muscle markers (desmin, myosin, dystrophin) was performed to characterize both the myoblasts and myotubes derived from each cell type and to test the transdifferentiation-inducing capacity of MYOD1 in fibroblasts and USCs. Specifically, the Duchenne muscular dystrophy (DMD) transcripts and proteins, including both the full-length Dp427 and the short Dp71 isoform, were evaluated. The protocol was firstly developed in healthy donor fibroblasts and USCs and then used to convert DMD patients' fibroblasts, with the aim of testing the efficacy of an antisense drug in vitro. Technical issues, limitations, and problems are explained and discussed. We demonstrate that MyoD-induced-fibroblasts and USCs are a useful in vitro model of myogenic cells to investigate possible therapies for neuromuscular diseases.
| Type: | Article |
|---|---|
| Title: | MyoD-induced reprogramming of human fibroblasts and urinary stem cells in vitro: protocols and their applications |
| Location: | Switzerland |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.3389/fphys.2023.1145047 |
| Publisher version: | https://doi.org/10.3389/fphys.2023.1145047 |
| Language: | English |
| Additional information: | © 2023 Rossi, Torelli, Ala, Weston, Morgan, Malhotra and Muntoni. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
| Keywords: | Duchenne muscular dystrophy, MyoD-induction, reprogramming, fibroblasts, urinary stem cells, lentivirus |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Population Health Sciences > UCL GOS Institute of Child Health > Developmental Neurosciences Dept |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10171857 |
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