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Time‐resolved video analysis and management system for monitoring cardiomyocyte differentiation processes and toxicology assays
Author(s) -
Ting Sherwin,
Liew Seaw Jia,
Japson Francis,
Shang Fuchun,
Chong Wee Keat,
Reuveny Shaul,
Tham Jo Yew,
Li Xiang,
Oh Steve
Publication year - 2014
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201300262
Subject(s) - embryoid body , myosin , cellular differentiation , embryonic stem cell , biology , biomedical engineering , microbiology and biotechnology , computational biology , medicine , induced pluripotent stem cell , biochemistry , gene
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.