Generation of craniofacial myogenic progenitor cells from human induced pluripotent stem cells for skeletal muscle tissue regeneration

• Type of Material: Article
• Authors: Eunhye Kim, Emory University; Fang Wu, Emory University; Xuewen Wu, Emory University; Hyojung Choo, Emory University
• Language: English
• Date: 2020-07-01
• Publisher: ELSEVIER SCI LTD
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2020 The Authors. Published by Elsevier Ltd.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.biomaterials.2020.119995
• Title of Journal or Parent Work: BIOMATERIALS
• Volume: 248
• Start Page: 119995
• End Page: 119995
• Grant/Funding Information: This research was supported in part by grants from National Institutes of Arthritis and Musculoskeletal and Skin Disease, United States, R01 AR071397 and National Research Foundation of Korea, South Korea, NRF-2018R1A6A3A03011703.
• Keywords: SATELLITE CELLS; DERIVATION; EXTRAOCULAR-MUSCLE; MESODERM; Technology; Muscle tissue engineering; Craniofacial myogenesis; POPULATION; Engineering; EXPANSION; Materials Science; Materials Science, Biomaterials; Craniofacial myogenic precursor cells; Science & Technology; PURIFICATION; EXPRESSION; SPECIFICATION; Engineering, Biomedical; Direct differentiation; MUSCULAR-DYSTROPHY; Human induced pluripotent stem cells

Abstract

Craniofacial skeletal muscle is composed of approximately 60 muscles, which have critical functions including food uptake, eye movements and facial expressions. Although craniofacial muscles have significantly different embryonic origin, most current skeletal muscle differentiation protocols using human induced pluripotent stem cells (iPSCs) are based on somite-derived limb and trunk muscle developmental pathways. Since the lack of a protocol for craniofacial muscles is a significant gap in the iPSC-derived muscle field, we have developed an optimized protocol to generate craniofacial myogenic precursor cells (cMPCs) from human iPSCs by mimicking key signaling pathways during craniofacial embryonic myogenesis. At each different stage, human iPSC-derived cMPCs mirror the transcription factor expression profiles seen in their counterparts during embryo development. After the bi-potential cranial pharyngeal mesoderm is established, cells are committed to cranial skeletal muscle lineages with inhibition of cardiac lineages and are purified by flow cytometry. Furthermore, identities of iPSC-derived cMPCs are verified with human primary myoblasts from craniofacial muscles using RNA sequencing. These data suggest that our new method could provide not only in vitro research tools to study muscle specificity of muscular dystrophy but also abundant and reliable cellular resources for tissue engineering to support craniofacial reconstruction surgery.