Enhanced Development of Skeletal Myotubes from Porcine Induced Pluripotent Stem Cells.
The pig is recognized as a valuable model in biomedical research in addition to its agricultural importance. Here we describe a means for generating skeletal muscle efficiently from porcine induced pluripotent stem cells (piPSC) in vitro thereby providing a versatile platform for applications ranging from regenerative biology to the ex vivo cultivation of meat. The GSK3B inhibitor, CHIR99021 was employed to suppress apoptosis, elicit WNT signaling events and drive naïve-type piPSC along the mesoderm lineage, and, in combination with the DNA methylation inhibitor 5-aza-cytidine, to activate an early skeletal muscle transcription program. Terminal differentiation was then induced by activation of an ectopically expressed MYOD1. Myotubes, characterized by myofibril development and both spontaneous and stimuli-elicited excitation-contraction coupling cycles appeared within 11 days. Efficient lineage-specific differentiation was confirmed by uniform NCAM1 and myosin heavy chain expression. These results provide an approach for generating skeletal muscle that is potentially applicable to other pluripotent cell lines and to generating other forms of muscle.
C.S. Bond Life Sciences Center, University of Missouri, Columbia, MO 6521, USA.
Nicholas J
NJ
Genovese
Department of Medical Pharmacology and Physiology, University of Missouri, Columbia, MO 65211, USA.
Timothy L
TL
Domeier
Department of Animal and Avian Sciences, University of Maryland, College Park, MD 20742, USA.|Animal Bioscience and Biotechnology Laboratory, USDA ARS, Beltsville, MD 20705, USA.
Bhanu Prakash V L
BP
Telugu
C.S. Bond Life Sciences Center, University of Missouri, Columbia, MO 6521, USA.
R Michael
RM
Roberts
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology* / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D013552:Swine; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism
C473711:Chir 99021; D011725:Pyridines; D011743:Pyrimidines; D051176:beta Catenin