title: Time-resolved video analysis and management system for monitoring cardiomyocyte differentiation processes and toxicology assays.
abstract: Cardiomyocytes (CM) derived from human embryonic stem cells (hESC) are used for cardio-toxicity evaluation and tested in many preclinical trials for their potential use in regenerative therapeutics. As more efficient CM differentiation protocols are developed, reliable automated platforms for characterization and detection are needed. An automated time-resolved video analysis and management system (TVAMS) has been developed for the evaluation of hESC differentiation to CM. The system was used for monitoring the kinetics of embryoid bodies (EB) generation (numbers and size) and differentiation into beating EBs (percentage beating area and beating EB count) in two differentiation protocols. We show that the percentage beating areas of EBs (from total area of the EBs) is a more sensitive and better predictor of CM differentiation efficiency than percentage of beating EBs (from total EBs) as the percentage beating areas of EBs correlates with cardiac troponin-T and myosin heavy chain expression levels. TVAMS can also be used to evaluate the effect of drugs and inhibitors (e.g. isoproterenol and ZD7288) on CM beating frequency. TVAMS can reliably replace the commonly practiced, time consuming, manual counting of total and beating EBs during CM differentiation. TVAMS is a high-throughput non-invasive video imaging platform that can be applied for the development of new CM differentiation protocols, as well as a tool to conduct CM toxicology assays.
authors.affiliation: Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), Singapore.
authors.forename: Sherwin
authors.initials: S
authors.lastname: Ting
authors.forename: Seaw Jia
authors.initials: SJ
authors.lastname: Liew
authors.forename: Francis
authors.initials: F
authors.lastname: Japson
authors.forename: Fuchun
authors.initials: F
authors.lastname: Shang
authors.forename: Wee Keat
authors.initials: WK
authors.lastname: Chong
authors.forename: Shaul
authors.initials: S
authors.lastname: Reuveny
authors.forename: Jo Yew
authors.initials: JY
authors.lastname: Tham
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Steve
authors.initials: S
authors.lastname: Oh
mesh_terms: D057005:Bioengineering / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D058732:Embryoid Bodies / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology; D059008:Time-Lapse Imaging / Q000379:methods*; D014741:Video Recording / Q000379:methods*
keywords: Bioprocess engineering; Imaging; Process monitoring; Stem cells; Toxicology