Development of a skeletal muscle sheet with direct reprogramming-induced myoblasts on a nanogel-cross-linked porous freeze-dried gel scaffold in a mouse gastroschisis model

• Published in: Pediatric surgery international Vol. 40; no. 1; p. 241
• Main Authors: Nagano, Shinta, Fumino, Shigehisa, Kishida, Tsunao, Wakao, Junko, Hirohata, Yoshiaki, Takayama, Shohei, Kim, Kiyokazu, Akiyoshi, Kazunari, Mazda, Osam, Tajiri, Tatsuro, Ono, Shigeru
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 26-08-2024; Springer Nature B.V
• Subjects: Animals; Disease Models, Animal; Freeze Drying - methods; Gastroschisis - surgery; Humans; Medicine; Medicine & Public Health; Mice; Mice, SCID; Muscle, Skeletal; Musculoskeletal system; Myoblasts; Nanogels; Original Article; Pediatric Surgery; Pediatrics; Polyethylene Glycols; Polyethyleneimine; Porosity; Surgery; Tissue Engineering - methods; Tissue Scaffolds; Myoblast; Direct reprogramming; Gastroschisis; Nanogel; Biosheet
• ISSN: 0179-0358; 1437-9813; 1437-9813
• DOI: 10.1007/s00383-024-05811-z

Purpose
In this study, we attempted to create skeletal muscle sheets made of directly converted myoblasts (dMBs) with a nanogel scaffold on a biosheet using a mouse gastroschisis model.
Methods
dMBs were prepared by the co-transfection of
MYOD1
and
MYCL
into human fibroblasts. Silicon tubes were implanted under the skin of NOG/SCID mice, and biosheets were formed. The nanogel was a nanoscale hydrogel based on cholesterol-modified pullulan, and a NanoClip-FD gel was prepared by freeze-drying the nanogel. 7 mm in length was created in the abdominal wall of NOG/SCID mice as a mouse gastroschisis model. Matrigel or NanoCliP-FD gel seeded with dMBs was placed on the biosheet and implanted on the model mice.
Results
Fourteen days after surgery, dMBs with Matrigel showed a small amount of coarse aggregations of muscle-like cells. In contrast, dMBs with NanoCliP-FD gel showed multinucleated muscle-like cells, which were expressed as desmin and myogenin by fluorescent immunostaining.
Conclusion
Nanogels have a porous structure and are useful as scaffolds for tissue regeneration by supplying oxygen and nutrients supply to the cells. Combining dMBs and nanogels on the biosheets resulted in the differentiation and engraftment of skeletal muscle, suggesting the possibility of developing skeletal muscle sheets derived from autologous cells and tissues.