title: Elastic hydrogel substrate supports robust expansion of murine myoblasts and enhances their engraftment.
abstract: 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 of 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.
authors.affiliation: Department of Pediatric Surgery, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610072, China. Electronic address: dk1118@yeah.net.
authors.forename: Ke
authors.initials: K
authors.lastname: Ding
authors.affiliation: Department of Clinical Hematology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Department of Orthopaedics, Southwest Hospital, Third Military Medical University, Chongqing 400038, China. Electronic address: xjzspine@163.com.
authors.forename: Jian-Zhong
authors.initials: JZ
authors.lastname: Xu
authors.affiliation: Department of Pediatric Surgery, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610072, China.
authors.forename: Wen-Ying
authors.initials: WY
authors.lastname: Liu
authors.affiliation: Department of Pediatric Surgery, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610072, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zeng
authors.affiliation: Department of Pediatric Surgery, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu 610072, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Hou
authors.affiliation: Department of Anatomy and Histology & Embryology, Chengdu Medical College, Chengdu 610500, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Lin
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D004548:Elasticity; D020100:Hydrogels / Q000737:chemistry*; D008810:Mice, Inbred C57BL; D008819:Mice, Nude; D008822:Mice, Transgenic; D032446:Myoblasts / Q000502:physiology* / Q000637:transplantation; D023822:Tissue Engineering; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D000943:Antigens, Differentiation; D020100:Hydrogels
keywords: Cell culture; Cell therapy; Cell transplantation; Elasticity; Myoblast; Stemness