Preparation of collagen-coated gels that maximize in vitro myogenesis of stem cells by matching the lateral elasticity of in vivo muscle.
The physical nature of a cell's microenvironment--including the elasticity of the surrounding tissue--appears to exert a significant influence on cell morphology, cytoskeleton, and gene expression. We have previously shown that committed muscle cells will develop sarcomeric striations of skeletal muscle myosin II only when the cells are grown on a compliant gel that closely matches the passive compliance of skeletal muscle. We have more recently shown with the same types of elastic gels that mesenchymal stem cells (MSCs) maximally express myogenic genes, even in the absence of tailored soluble factors. Here, we provide detailed methods not only for how we make and nanomechanically characterize hydrogels of muscle-like elasticity, but also how we culture MSCs and characterize their myogenic induction by whole human genome transcript analysis.
Chaudhuri
Tathagata
T
Pennsylvania Muscle Institute, University of Pennsylvania, Philadelphia, PA, USA.
Rehfeldt
Florian
F
Sweeney
H Lee
HL
Discher
Dennis E
DE
D000180:Acrylic Resins; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D003094:Collagen / Q000378:metabolism*; D055119:Elastic Modulus; D004548:Elasticity; D005455:Fluorescent Antibody Technique; D005782:Gels; D005898:Glass; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D018625:Microscopy, Atomic Force; D024510:Muscle Development; D009132:Muscles / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D011506:Proteins / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D000099166:Polyacrylamides
D000180:Acrylic Resins; D015415:Biomarkers; D003094:Collagen; D005782:Gels; D011506:Proteins; D012333:RNA, Messenger; D000099166:Polyacrylamides
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