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Title: Elastic hydrogel substrate supports robust expansion of murine myoblasts and enhances their engraftment

The application of satellite cell-derived myoblasts in regenerative medicine has been restricted by the rapid loss of stemness during in vitro cell expansion using traditional culture systems. However, studies published in the past decade have highlighted the influence of substrate elasticity on stem cell fate and revealed that culture on a soft hydrogel substrate can promote self-renewal and prolong the regenerative potential of muscle stem cells. Whether hydrogel substrates have similar effects after long-term robust expansion remains to be determined. Herein we prepared an elastic chitosan/beta-glycerophosphate/collagen hydrogel mimicking the soft microenvironment of muscle tissues for use as the substrate for satellite cell culture and investigated its influence on long-term cell expansion. After 20 passages in culture, satellite cell-derived myoblasts cultured on our hydrogel substrate exhibited significant improvements in proliferation capability, cell viability, colony forming frequency, and potential for myogenic differentiation compared to those cultured on a routine rigid culture surface. Immunochemical staining and western blot analysis both confirmed that myoblasts cultured on the hydrogel substrate expressed higher levels of several differentiation-related markers, including Pax7, Pax3, and SSEA-1, and a lower level of MyoD compared to myoblasts cultured on rigid culture plates (all p<0.05). After transplantation into the tibialis anterior ofmore » nude mice, myoblasts that had been cultured on the hydrogel substrate demonstrated a significantly greater engraftment efficacy than those cultured on the traditional surface. Collectively, these results indicate that the elastic hydrogel substrate supported robust expansion of murine myoblasts and enhanced their engraftment in vivo. - Highlights: • An elastic hydrogel was designed to mimic the pliable muscle tissue microenvironment. • Myoblasts retained their stemness in long-term culture on the elastic hydrogels. • Myoblasts expanded on the elastic hydrogel exhibited enhanced in vivo engraftment.« less
Authors:
 [1] ;  [2] ;  [3] ; ; ;  [1] ;  [4]
  1. Department of Pediatric Surgery, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Chengdu 610072 (China)
  2. Department of Clinical Hematology, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China)
  3. Department of Orthopaedics, Southwest Hospital, Third Military Medical University, Chongqing 400038 (China)
  4. Department of Anatomy and Histology & Embryology, Chengdu Medical College, Chengdu 610500 (China)
Publication Date:
OSTI Identifier:
22462330
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 337; Journal Issue: 1; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AMINO ACIDS; ANIMAL TISSUES; CELL CULTURES; COLLAGEN; DRUGS; EXPANSION; HYDROGELS; IN VITRO; IN VIVO; MICE; MYOBLASTS; OLIGOSACCHARIDES; STEM CELLS; SUBSTRATES; THERAPY