title: Using image-based flow cytometry to monitor satellite cells proliferation and differentiation in vitro.
abstract: Muscle satellite cells can proliferate and differentiate into myocytes; this biological function has important implications for muscle development, aging, repair, and wasting diseases. Established analytical methods, including western blotting, PCR, and immunohistochemistry, has been used to characterize the stages of satellite cells growth and development but there is currently a lack of methods to simultaneously monitor cell proliferation and identify changes in cell properties (i.e., expression of myogenic regulatory factors) for each individual cell during the progression to become myocytes. Image-based flow cytometry has the capability to simultaneously obtain morphometric data, monitor cell division, and detect expression of multiple proteins of interest. In this article we demonstrate the use of image-based flow cytometry and cell-trace dye to monitor satellite cells proliferation and to identify different stages of satellite cell differentiation.
authors.affiliation: Applied Physiology Laboratory, Department of Kinesiology, Health Promotion, and Recreation, University of North Texas, 1155 Union Circle #310769, Denton, TX 76203-5017, United States; Department of Biological Sciences, University of North Texas, 1155 Union Circle #305220, Denton, TX 76203-5017, United States.
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Luk
authors.affiliation: Applied Physiology Laboratory, Department of Kinesiology, Health Promotion, and Recreation, University of North Texas, 1155 Union Circle #310769, Denton, TX 76203-5017, United States; Department of Biological Sciences, University of North Texas, 1155 Union Circle #305220, Denton, TX 76203-5017, United States.
authors.forename: Brian K
authors.initials: BK
authors.lastname: McFarlin
authors.affiliation: Applied Physiology Laboratory, Department of Kinesiology, Health Promotion, and Recreation, University of North Texas, 1155 Union Circle #310769, Denton, TX 76203-5017, United States; Department of Biological Sciences, University of North Texas, 1155 Union Circle #305220, Denton, TX 76203-5017, United States. Electronic address: Jakob.Vingren@unt.edu.
authors.forename: Jakob L
authors.initials: JL
authors.lastname: Vingren
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D049109:Cell Proliferation; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D032342:Muscle Cells / Q000166:cytology* / Q000378:metabolism; D017570:MyoD Protein / Q000235:genetics / Q000378:metabolism; D051763:PAX7 Transcription Factor / Q000235:genetics / Q000378:metabolism; D061251:Primary Cell Culture; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D013388:Succinimides / Q000737:chemistry
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D015415:Biomarkers; D005452:Fluoresceins; D005456:Fluorescent Dyes; D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; D051763:PAX7 Transcription Factor; C497321:PAX7 protein, human; D013388:Succinimides
keywords: Differentiation; MyoD; Myoblasts; Myocytes; Pax7; Proliferation