Time-resolved video analysis and management system for monitoring cardiomyocyte differentiation processes and toxicology assays
Time-resolved video analysis and management system for monitoring cardiomyocyte differentiation processes and toxicology assays
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DOI：
10.1002/biot.201300262
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发表时间：
2014-05-01
影响因子：
4.7
通讯作者：
Oh, Steve
中科院分区：
文献类型：
--
作者：
Ting, Sherwin;Liew, Seaw Jia;Oh, Steve
来源于人胚胎干细胞（hESC）的心肌细胞（CM）用于心脏毒性评价，并在许多临床前试验中测试其在再生治疗中的潜在用途。随着更有效的CM分化方案的开发，需要用于表征和检测的可靠的自动化平台。一个自动化的时间分辨视频分析和管理系统（TVAMS）已经开发用于评估人胚胎干细胞分化为CM。该系统用于监测两种分化方案中胚状体（EB）产生（数量和大小）和分化成搏动EB（搏动面积百分比和搏动EB计数）的动力学。我们发现，EBs的搏动面积百分比（来自EBs的总面积）比搏动EBs的百分比（来自总EBs）更敏感，更好地预测CM分化效率，因为EBs的搏动面积百分比与心肌肌钙蛋白-T和肌球蛋白重链表达水平相关。TVAMS也可用于评价药物和抑制剂（如异丙肾上腺素和ZD 7288）对CM搏动频率的影响。TVAMS可以可靠地取代CM分化过程中常见的耗时的总EB和跳动EB的手动计数。TVAMS是一种高通量非侵入性视频成像平台，可用于开发新的CM分化方案，以及进行CM毒理学测定的工具。
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.