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Addressing the scalability of human iPSC-derived neurons for HTS implementation and phenotypic screening

Speaker

Abstract

Traditional high throughput screening (HTS) assays for neuronal targets employ non-human primary neuronal cells due to the scale necessary for HTS. Isolation of mouse primary neurons can be unreliable and economically demanding. The discovery of new drugs for neuropsychiatric disorders has further been hampered by lack of access to disease-relevant human primary neurons and appropriate disease models. By using human induced pluripotent stem cell (hiPSC) technology, we can address some of the obstacles which affords the generation of human neurons through (1) embryoid body (EB) formation, (2) cultivation on stromal feeder cells, and, (3) employing lineage specific differentiation factors. Methods exist to reproducibly differentiate hiPSCs into functional cortical induced neurons (iN) in less than two weeks but, they have never been taken to the HTS scale and have been slow and somewhat variable. We have successfully recapitulated the aforementioned technique and leveraged the CRISPR technology to define the path to a plate-compatible format amenable for large-scale HTS implementation. The resulting iN cells exhibit appropriate genetic and fluorescent markers that give confidence of bonafide neuronal differentiation. Imminently, we intent to test the preliminary iN cells for their ability to post-mitotically increase or decrease neurite outgrowth following treatment with LOPAC test compounds via staining and high content analysis. Ultimately, we will determine reliability and reproducibility over time with industrial scale robotics. Furthermore, we also intend to leverage the CRISPR technology to create a library of disease-relevant-phenotypes from hiPSC-derived cellular models that will provide more opportunities for all biologists to study epigenetic mechanisms and scale-up screening initiatives with Scripps Research Institute Molecular Screening Center (SRIMSC).


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Addressing the scalability of human iPSC-derived neurons for HTS implementation and phenotypic screening


Specialty

Animal Diets

Drug Discovery

Crispr

Chemistry

Research

Cancer Therapeutics

Biology

Biochemistry

Molecular Biology

Animal Models

Gene Expression

Crispr-Cas9

Clinical Chemistry

Cell Biology

Animal Research

Geography

North America100%

Registration Source

Website Visitors100%

Job Title

Research Scientist100%

Organization

Research Institute100%


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