scholarly journals Multimodal on-axis platform for all-optical electrophysiology with near-infrared probes in human stem-cell-derived cardiomyocytes

2018 ◽  
Author(s):  
Aleksandra Klimas ◽  
Gloria Ortiz ◽  
Steven Boggess ◽  
Evan W. Miller ◽  
Emilia Entcheva
Keyword(s):  
Stem Cell ◽  
High Throughput ◽  
High Performance ◽  
Near Infrared ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Simultaneous Recording ◽  
Optogenetic Stimulation ◽  
All Optical ◽  
Induced Pluripotent

AbstractCombined optogenetic stimulation and optical imaging permits scalable, high-throughput probing of cellular electrophysiology and optimization of stem-cell derived excitable cells, such as neurons and muscle cells. We report a new “on-axis” configuration of OptoDyCE, our all-optical platform for studying human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) and other cell types, optically driven by Channelrhodopsin2 (ChR2). This solid-state system integrates optogenetic stimulation with temporally-multiplexed simultaneous recording of membrane voltage (Vm) and intracellular calcium ([Ca2+]i) dynamics using a single photodetector. We demonstrate the capacity for combining multiple spectrally-compatible actuators and sensors, including newer high-performance near-infrared (NIR) voltage probes BeRST1 and Di-4-ANBDQBS, to record complex spatiotemporal responses of hiPSC-CMs to drugs in a high-throughput manner.

High-throughput screening of human induced pluripotent stem cell-derived brain organoids

2020 ◽  
Vol 335 ◽  
pp. 108627 ◽  
Author(s):  
Madel Durens ◽  
Jonathan Nestor ◽  
Madeline Williams ◽  
Kevin Herold ◽  
Robert F. Niescier ◽  
...  
Keyword(s):  
Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Induced Pluripotent

scholarly journals A high-throughput screening platform for pigment regulating agents using pluripotent stem cell derived melanocytes

2020 ◽  
Author(s):  
Valentin Parat ◽  
Brigitte Onteniente ◽  
Julien Maruotti
Keyword(s):  
Stem Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Light Scatter ◽  
Chemical Treatments ◽  
Human Melanocytes ◽  
Screening Platform ◽  
Induced Pluripotent

AbstractIn this study, we describe a simple and straight-forward assay using induced pluripotent stem cell derived melanocytes and high-throughput flow cytometry, to screen and identify pigment regulating agents. The assays is based on the correlation between forward light-scatter characteristics and melanin content, with pigmented cells displaying high light absorption/low forward light-scatter, while the opposite is true for lowly pigmented melanocytes, as a result of genetic background or chemical treatments. Orthogonal validation is then performed by regular melanin quantification. Such approach was validated using a set of 80 small molecules, and yielded a confirmed hit. The assay described in this study may prove a useful tool to identify modulators of melanogenesis in human melanocytes.

scholarly journals A new platform for high-throughput therapy testing on iPSC-derived, immature airway from Cystic Fibrosis Patients

2021 ◽  
Author(s):  
Jia Xin Jiang ◽  
Leigh Wellhauser ◽  
Onofrio Laselva ◽  
Irina Utkina ◽  
Zoltan Bozoky ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Stem Cell ◽  
High Throughput ◽  
Induced Pluripotent Stem Cell ◽  
Patient Specific ◽  
Functional Responses ◽  
Primary Defect ◽  
Epithelial Cultures ◽  
Therapy Development ◽  
Induced Pluripotent

Induced pluripotent, stem cell (iPSC)-derived models of airway tissue have successfully modeled the primary defect in regulated chloride conductance caused by the major Cystic Fibrosis causing mutation, F508del. However, it remains unclear if iPSC-derived airway cultures can be used in high-throughput therapy development for F508del and rarer mutations. There is an urgent need for airway tissue models that reflect the variability of patient-specific responses and are scalable for therapy development. In the current work, we describe a robust, high-throughput fluorescence assay of mutant CFTR function in iPSCs differentiated to immature airway epithelium. This assay measures reproducible functional responses to modulators targeting either the major CF mutant F508del or the nonsense mutant: W1282X-CFTR. We show that the ranking of patient-specific responses to interventions in this stem-cell based model recapitulates the ranking observed in primary nasal epithelial cultures obtained from the same individuals. In summary, these proof-of-concept studies show that this novel platform has the potential to support therapy development and precision medicine for Cystic Fibrosis patients.

scholarly journals Induced pluripotent stem cell technology for modelling and therapy of cerebellar ataxia

Open Biology ◽  
2015 ◽  
Vol 5 (7) ◽  
pp. 150056 ◽  
Author(s):  
Lauren M. Watson ◽  
Maggie M. K. Wong ◽  
Esther B. E. Becker
Keyword(s):  
Stem Cell ◽  
Cerebellar Ataxia ◽  
Pluripotent Stem Cell ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Disease Modelling ◽  
Stem Cell Technology ◽  
Induced Pluripotent ◽  
First Descriptions ◽  
Made In

Induced pluripotent stem cell (iPSC) technology has emerged as an important tool in understanding, and potentially reversing, disease pathology. This is particularly true in the case of neurodegenerative diseases, in which the affected cell types are not readily accessible for study. Since the first descriptions of iPSC-based disease modelling, considerable advances have been made in understanding the aetiology and progression of a diverse array of neurodegenerative conditions, including Parkinson's disease and Alzheimer's disease. To date, however, relatively few studies have succeeded in using iPSCs to model the neurodegeneration observed in cerebellar ataxia. Given the distinct neurodevelopmental phenotypes associated with certain types of ataxia, iPSC-based models are likely to provide significant insights, not only into disease progression, but also to the development of early-intervention therapies. In this review, we describe the existing iPSC-based disease models of this heterogeneous group of conditions and explore the challenges associated with generating cerebellar neurons from iPSCs, which have thus far hindered the expansion of this research.

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