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J Biomol Screen. 2016 Dec;21(10):1112-1124. doi: 10.1177/1087057116670068. Epub 2016 Sep 26.

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays.

Author information

1
1 National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
2
2 Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China.
3
3 Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.

Abstract

Human neuronal cells differentiated from induced pluripotent cells have emerged as a new model system for the study of disease pathophysiology and evaluation of drug efficacy. Differentiated neuronal cells are more similar in genetics and biological content to human brain cells than other animal disease models. However, culture of neuronal cells in assay plates requires a labor-intensive procedure of plate precoating, hampering its applications in high-throughput screening (HTS). We developed a simplified method with one-step seeding of neural stem cells in assay plates by supplementing the medium with a recombinant human vitronectin (VTN), thus avoiding plate precoating. Robust results were obtained from cell viability, calcium response, and neurite outgrowth assays using this new method. Our data demonstrate that this approach greatly simplifies high-throughput assays using neuronal cells differentiated from human stem cells for translational research.

KEYWORDS:

HTS; high-content screening; high-throughput screening; iPSC; induced pluripotent stem cells; neural stem cells; neuronal cells; plate coating; vitronectin

PMID:
27647668
PMCID:
PMC5285267
DOI:
10.1177/1087057116670068
[Indexed for MEDLINE]
Free PMC Article

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