title: Real-time quantitative monitoring of hiPSC-based model of macular degeneration on Electric Cell-substrate Impedance Sensing microelectrodes.
abstract: 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 Ω(1/2)cm for the case cell line and 6.5 ± 0.15 Ω(1/2)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.
authors.affiliation: Institute for Bioengineering, School of Engineering, The University of Edinburgh, EH9 3DW, United Kingdom.
authors.forename: W
authors.initials: W
authors.lastname: Gamal
authors.affiliation: MRC Centre for Regenerative Medicine, The University of Edinburgh, EH16 4UU, United Kingdom; Centre for Clinical Brain Sciences, The University of Edinburgh, EH16 4SB, United Kingdom; Euan MacDonald Centre for MND Research, The University of Edinburgh, EH16 4SB, United Kingdom; Centre for Neuroregeneration, The University of Edinburgh, EH16 4SB, United Kingdom; The Anne Rowling Regenerative Neurology Clinic, The University of Edinburgh, EH16 4SB, United Kingdom.
authors.forename: S
authors.initials: S
authors.lastname: Borooah
authors.affiliation: Institute for Bioengineering, School of Engineering, The University of Edinburgh, EH9 3DW, United Kingdom.
authors.forename: S
authors.initials: S
authors.lastname: Smith
authors.affiliation: Institute for Integrated Micro and Nano Systems, School of Engineering, The University of Edinburgh, EH9 3JF, United Kingdom.
authors.forename: I
authors.initials: I
authors.lastname: Underwood
authors.affiliation: Institute for Bioengineering, School of Engineering, The University of Edinburgh, EH9 3DW, United Kingdom.
authors.forename: V
authors.initials: V
authors.lastname: Srsen
authors.affiliation: MRC Centre for Regenerative Medicine, The University of Edinburgh, EH16 4UU, United Kingdom; Centre for Clinical Brain Sciences, The University of Edinburgh, EH16 4SB, United Kingdom; Euan MacDonald Centre for MND Research, The University of Edinburgh, EH16 4SB, United Kingdom; Centre for Neuroregeneration, The University of Edinburgh, EH16 4SB, United Kingdom; The Anne Rowling Regenerative Neurology Clinic, The University of Edinburgh, EH16 4SB, United Kingdom.
authors.forename: S
authors.initials: S
authors.lastname: Chandran
authors.affiliation: MRC Centre for Regenerative Medicine, The University of Edinburgh, EH16 4UU, United Kingdom. Electronic address: Pierre.Bagnaninchi@ed.ac.uk.
authors.forename: P O
authors.initials: PO
authors.lastname: Bagnaninchi
authors.affiliation: Centre for Clinical Brain Sciences, The University of Edinburgh, EH16 4SB, United Kingdom; The Anne Rowling Regenerative Neurology Clinic, The University of Edinburgh, EH16 4SB, United Kingdom; School of Clinical Sciences, The University of Edinburgh, EH16 4SB, United Kingdom.
authors.forename: B
authors.initials: B
authors.lastname: Dhillon
mesh_terms: D001681:Biological Assay / Q000295:instrumentation*; D002465:Cell Movement; D002478:Cells, Cultured; D003199:Computer Systems; D058266:Dielectric Spectroscopy / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008268:Macular Degeneration / Q000473:pathology* / Q000503:physiopathology; D008839:Microelectrodes; D015203:Reproducibility of Results; D055213:Retinal Pigment Epithelium / Q000473:pathology* / Q000502:physiology; D012680:Sensitivity and Specificity; D046888:Tissue Array Analysis / Q000295:instrumentation*
keywords: Disease model; Human induced pluripotent stem cells; Impedance sensing; Macular degeneration; Tissue-on-a-chip; Wound healing