Non-invasive characterization of mouse embryonic stem cell derived cardiomyocytes based on the intensity variation in digital beating video
Non-invasive characterization of mouse embryonic stem cell derived cardiomyocytes based on the intensity variation in digital beating video
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DOI：
10.1039/c0an00208a
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发表时间：
2010-01-01
期刊：
影响因子：
4.2
通讯作者：
Tamiya, Eiichi
中科院分区：
文献类型：
--
作者：
Hossain, Mohammad Mosharraf;Shimizu, Eiichi;Tamiya, Eiichi
由于胚胎干（ES）细胞或诱导多能干（iPS）细胞分化产生的心肌细胞具有再生治疗和作为药物筛选的药物模型的潜力，人们对它们的兴趣日益增加。 ES 或 iPS 衍生的心肌细胞的表征对于此类细胞的预期用途而言具有挑战性且不可避免。在本文中，我们概述了一种评估心肌细胞体外跳动特性的新型非侵入性方法。该方法基于对从搏动组织的光学显微视频记录的连续收缩帧和舒张帧获得的导数图像中总像素强度随时间变化的分析。快速傅立叶变换 (FFT) 生成这些图像的频域。分析的信噪比符合罗斯标准。我们已成功应用我们的方法监测小鼠 ES 细胞 (mESC) 衍生的心肌细胞，以确定搏动的起始、搏动组织的组织和成熟，根据搏动间隔或频率以及搏动强度计算搏动节律。我们已经展示了我们的图像分析方法在直接监测分化心肌细胞对水合咖啡因、对羟基苯乙酰胺和脱水氯化钙（分别作为正性、中性和负性肌力药物）的反应方面的成功应用。这种非侵入性的表征方法将有助于研究这些细胞对各种外部刺激（例如分化促进剂或治疗）的反应，以及以快速且廉价的方式进行初步药物筛选，而无需太多专业知识。
The interest in cardiomyocytes derived from differentiation of embryonic stem (ES) cells or induced pluripotent stem (iPS) cells is increasing due to their potential for regenerative therapeutics and as a pharmaceutical model of drug screening. Characterization of ES or iPS derived cardiomyocytes is challenging and inevitable for the intended usage of such cells. In this paper we have outlined a novel, non-invasive method for evaluating in vitro beating properties of cardiomyocytes. The method is based on the analysis of time dependent variation in the total pixel intensities in derivative images obtained from the consecutive systolic and diastolic frames from the light microscopic video recordings of beating tissue. Fast Fourier transform (FFT) yielded the frequency domains for these images. The signal to noise ratio for the analysis met the Rose criterion. We have successfully applied our method for monitoring mouse ES cell (mESC) derived cardiac muscle cells to determine the initiation of beating, organization and maturation of beating tissue, calculating the beating rhythms in terms of beating interval or frequency and the strength of beating. We have shown the successful application of our image analysis method in direct monitoring of the responses of differentiated cardiomyocytes towards caffeine hydrate, p-hydroxyphenylacetamide and calcium chloride dehydrate-respectively as positive, neutral and negative inotropic agents. This non-invasive method of characterization will be useful in studying the response of these cells to various external stimulations, such as differentiation promoting agents or treatments, as well as in preliminary drug screening in a quick and inexpensive manner without needing much expertise.