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Promoting differentiation of cultured myoblasts using biomimetic surfaces that present alpha-laminin-2 peptides
Authors:Francine Parker  Kathryn White  Siȏn Phillips  Michelle Peckham
Affiliation:1School of Molecular and Cellular Biology, Faculty of Biological Sciences, The University of Leeds, Leeds, LS2 9JT UK ;2Bioscience Centre, International Centre for Life, Orla Protein Technologies Ltd, Newcastle upon Tyne, NE1 4EP UK
Abstract:Traditionally, muscle cell lines are cultured on glass coverslips and differentiated to investigate myoblast fusion and differentiation. Efficient differentiation of myoblasts produces a dense network of myotubes with the correct organisation for contraction. Here we have tested the ability of artificially generated, precisely controlled peptide surfaces to enhance the efficiency of myoblast differentiation. We focused on specific short peptides from α-laminin-2 (IKVSV, VQLRNGFPYFSY and GLLFYMARINHA) as well as residues 15–155 from FGF1. We tested if these peptides in isolation, and/or in combination promoted muscle differentiation in culture, by promoting fusion and/or by improving sarcomere organisation. The majority of these peptides promoted fusion and differentiation in two different mouse myogenic cell lines and in primary human myoblasts. The additive effects of all four peptides gave the best results for both mouse cell lines tested, while primary human cell cultures differentiated equally well on most peptide surfaces tested. These data show that a mixture of short biomimetic peptides can reliably promote differentiation in mouse and human myoblasts.
Keywords:Myoblast   Myogenesis   Laminin   LAMA2   Peptides   Differentiation
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