Real-time quantitative monitoring of hiPSC-based model of macular degeneration on Electric Cell-substrate Impedance Sensing microelectrodes.
Real-time quantitative monitoring of hiPSC-based model of macular degeneration on Electric Cell-substrate Impedance Sensing microelectrodes.
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DOI：
10.1016/j.bios.2015.04.079
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发表时间：
2015-09-15
影响因子：
12.6
通讯作者：
Dhillon, B.
中科院分区：
文献类型：
--
作者：
Gamal, W.;Borooah, S.;Smith, S.;Underwood, I.;Srsen, V.;Chandran, S.;Bagnaninchi, P. O.;Dhillon, B.
关键词：
视网膜相关性黄斑变性（AMD）是发达国家致盲的主要原因。需要人源化疾病模型来开发针对目前无法治愈的AMD形式的新疗法。在这项工作中，开发了一种组织芯片方法，通过结合人类诱导多能干细胞，电细胞基质阻抗传感（ECIS）和可重复的电损伤测定来建模和定量研究AMD。使用从患有遗传性黄斑变性的患者和未受影响的同胞产生的视网膜色素上皮（RPE）细胞来测试模型平台，在所述模型平台上进行可再现的电创伤测定以对RPE损伤建模。首先，在25天内进行了稳健且可重复的实时定量监测，证明了微电极阵列上RPE层的建立和成熟。然后开始模拟AMD疾病中细胞损失的空间控制的RPE层损伤。细胞恢复后的迁移率（8.6±0.46 μm/h）与对照组（10.69±0.21 μm/h）相比，差异有显著性（P<0.01）。定量数据分析表明，这是由于对照细胞系中细胞-底物粘附较低而实现的。ECIS细胞-基质粘附参数（α）为7.8±0.28 Ω1/2 cm（病例细胞系）和6.5±0.15 Ω1/2 cm（对照细胞系）。这些发现证实了使用细胞粘附生化测定。开发的疾病模型芯片是一个强大的转化研究平台，具有相当大的潜力，通过实现实时，定量和可重复的患者特异性RPE细胞修复研究来研究新的疗法。建立hiPSC-RPE模型并在ECIS微电极阵列上表征。使用可再现的电伤口愈合测定来模拟RPE细胞损伤。病例组和对照组之间的迁移率存在显著差异。细胞通过阻抗波动反映的间质迁移恢复。病例细胞系显示出明显高于对照细胞系的粘附力。
Age-related macular degeneration (AMD) is the leading cause of blindness in the developed world. Humanized disease models are required to develop new therapies for currently incurable forms of AMD. In this work, a tissue-on-a-chip approach was developed through combining human induced pluripotent stem cells, Electric Cell–substrate Impedance Sensing (ECIS) and reproducible electrical wounding assays to model and quantitatively study AMD. Retinal Pigment Epithelium (RPE) cells generated from a patient with an inherited macular degeneration and from an unaffected sibling were used to test the model platform on which a reproducible electrical wounding assay was conducted to model RPE damage. First, a robust and reproducible real-time quantitative monitoring over a 25-day period demonstrated the establishment and maturation of RPE layers on the microelectrode arrays. A spatially controlled RPE layer damage that mimicked cell loss in AMD disease was then initiated. Post recovery, significant differences (P<0.01) in migration rates were found between case (8.6±0.46 μm/h) and control cell lines (10.69±0.21 μm/h). Quantitative data analysis suggested this was achieved due to lower cell–substrate adhesion in the control cell line. The ECIS cell–substrate adhesion parameter (α) was found to be 7.8±0.28 Ω1/2 cm for the case cell line and 6.5±0.15 Ω1/2 cm for the control. These findings were confirmed using cell adhesion biochemical assays. The developed disease model-on-a-chip is a powerful platform for translational studies with considerable potential to investigate novel therapies by enabling real-time, quantitative and reproducible patient-specific RPE cell repair studies. hiPSC-RPE model was established and characterised on ECIS microelectrode arrays. A reproducible electrical wound healing assay was used to mimic RPE cell damage. Significant differences in migration rates were found between case and control. Cells recovered through mesenchymal migration mirrored by impedance fluctuations. Case cell line showed significantly higher adhesion than the control cell line.
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影响因子：
3.9
作者：
Lee, Jiyeon;Wang, Yu-Lin;Lele, Tanmay P.
通讯作者：
Lele, Tanmay P.
影响因子：
24.3
作者：
Heijink, I. H.;Brandenburg, S. M.;van Oosterhout, A. J. M.
通讯作者：
van Oosterhout, A. J. M.
DOI：
10.1073/pnas.1202922109
发表时间：
2012-04-10
影响因子：
11.1
作者：
Bilican, Bilada;Serio, Andrea;Chandran, Siddharthan
通讯作者：
Chandran, Siddharthan
影响因子：
4.8
作者：
Han, Jingyan;Liu, Guoquan;Voyno-Yasenetskaya, Tatyana
通讯作者：
Voyno-Yasenetskaya, Tatyana
影响因子：
2.7
作者：
Keese, CR;Bhawe, K;Giaever, I
通讯作者：
Giaever, I