Generation of allogeneic and xenogeneic functional muscle stem cells for intramuscular transplantation
Ajda Lenardič, … , Christoph Handschin, Ori Bar-Nur
Ajda Lenardič, … , Christoph Handschin, Ori Bar-Nur
Published May 7, 2024
Citation Information: J Clin Invest. 2024;134(12):e166998. https://doi.org/10.1172/JCI166998.
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Research Article Muscle biology

Generation of allogeneic and xenogeneic functional muscle stem cells for intramuscular transplantation

Abstract

Satellite cells, the stem cells of skeletal muscle tissue, hold a remarkable regeneration capacity and therapeutic potential in regenerative medicine. However, low satellite cell yield from autologous or donor-derived muscles hinders the adoption of satellite cell transplantation for the treatment of muscle diseases, including Duchenne muscular dystrophy (DMD). To address this limitation, here we investigated whether satellite cells can be derived in allogeneic or xenogeneic animal hosts. First, injection of CRISPR/Cas9-corrected Dmdmdx mouse induced pluripotent stem cells (iPSCs) into mouse blastocysts carrying an ablation system of host satellite cells gave rise to intraspecies chimeras exclusively carrying iPSC-derived satellite cells. Furthermore, injection of genetically corrected DMD iPSCs into rat blastocysts resulted in the formation of interspecies rat-mouse chimeras harboring mouse satellite cells. Notably, iPSC-derived satellite cells or derivative myoblasts produced in intraspecies or interspecies chimeras restored dystrophin expression in DMD mice following intramuscular transplantation and contributed to the satellite cell pool. Collectively, this study demonstrates the feasibility of producing therapeutically competent stem cells across divergent animal species, raising the possibility of generating human muscle stem cells in large animals for regenerative medicine purposes.

Authors

Ajda Lenardič, Seraina A. Domenig, Joel Zvick, Nicola Bundschuh, Monika Tarnowska-Sengül, Regula Furrer, Falko Noé, Christine L. Trautmann, Adhideb Ghosh, Giada Bacchin, Pjeter Gjonlleshaj, Xhem Qabrati, Evi Masschelein, Katrien De Bock, Christoph Handschin, Ori Bar-Nur

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Figure 4

Chimera-derived muscle stem cells restore dystrophin expression in DMD mice.

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Chimera-derived muscle stem cells restore dystrophin expression in DMD m...
(A) Schematic representation depicting the strategy for intramuscular transplantation. A similar number of satellite cells or expanded myoblasts were transplanted from the same mouse. (B) A graph showing the number of Pax7-nGFP+ cells obtained from the specified animals. n = 4 animals for Pax7-nGFP mice and n = 6 animals for chimeras. Data are presented as mean ± SD. Statistical analysis was performed using a Student’s 2-tailed t test. (C) Representative immunostaining images of tibialis anterior (TA) muscle cross section of Dmdmdx-4Cv; Prkdcscid mice stained for dystrophin 4 weeks after transplantation with the indicated cell lines. Scale bars: 1 mm (top panel) and 100 μm (bottom panel). Sat. cells, satellite cells; myobl., myoblasts. (D) Quantification of the transplantation trial shown in C. n = 5 animals for Pax7-nGFP mice and n = 6 animals for chimeras. Each dot represents 1 recipient, with colors specifying cells derived from the same donor. The number of transplanted cells from each donor is shown in Supplemental Figure 5A. Data are presented as mean ± SD. Statistical analysis was performed using a 2-way ANOVA. **P ≤ 0.01; ****P ≤ 0.0001. (E) Representative immunostaining images for dystrophin and fiber typing in TA muscle cross sections of Dmdmdx-4Cv; Prkdcscid mice 4 weeks after transplantation with the specified mouse-derived cell lines. Scale bars: 1 mm (top) and 100 μm (bottom). (F) Representative images of TA muscle cross section from Dmdmdx-4Cv; Prkdcscid mice immunostained for the indicated markers 4 weeks after transplantation with the specified cell lines. Arrowheads point to colocalization of PAX7 expression and the Pax7-nGFP reporter in rare cells. Scale bars: 100 μm (top) and 25 μm (bottom). LUTs were individually adjusted.