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3D-Cultured Neural Stem Cell Microarrays on a Micropillar Chip for High-Throughput Developmental Neurotoxicology
Experimental Cell Research
  • Pranav Joshi, Cleveland State University
  • Kyeong Nam Yu, Cleveland State University
  • Soo Yeon Kang, Cleveland State University
  • Seok Joon Kwon, Rensselaer Polytechnic Institute
  • Paul S. Kwon, Rensselaer Polytechnic Institute
  • Jonathan S. Dordick, Rensselaer Polytechnic Institute
  • Chandrasekhar R. Kothapalli, Cleveland State University
  • Moo Yeal Lee, Cleveland State University
Document Type
Article
Publication Date
9-15-2018
Disciplines
Abstract

© 2018 Numerous chemicals including environmental toxicants and drugs have not been fully evaluated for developmental neurotoxicity. A key gap exists in the ability to predict accurately and robustly in vivo outcomes based on in vitro assays. This is particularly the case for predicting the toxicity of chemicals on the developing human brain. A critical need for such in vitro assays is choice of a suitable model cell type. To that end, we have performed high-throughput in vitro assessment of proliferation and differentiation of human neural stem cells (hNSCs). Conventional in vitro assays typically use immunofluorescence staining to quantify changes in cell morphology and expression of neural cell-specific biomarkers, which is often time-consuming and subject to variable specificities of available antibodies. To alleviate these limitations, we developed a miniaturized, three-dimensional (3D) hNSC culture with ReNcell VM on microarray chip platforms and established a high-throughput promoter-reporter assay system using recombinant lentiviruses on hNSC spheroids to assess cell viability, self-renewal, and differentiation. Optimum cell viability and spheroid formation of 3D ReNcell VM culture were observed on a micropillar chip over a period of 9 days in a mixture of 0.75% (w/v) alginate and 1 mg/mL growth factor reduced (GFR) Matrigel with 25 mM CaCl2 as a crosslinker for alginate. In addition, 3D ReNcell VM culture exhibited self-renewal and differentiation on the microarray chip platform, which was efficiently monitored by enhanced green fluorescent protein (EGFP) expression of four NSC-specific biomarkers including sex determining region Y-box 2 (SOX2), glial fibrillary acidic protein (GFAP), synapsin1, and myelin basic protein (MBP) with the promoter-reporter assay system.

DOI
10.1016/j.yexcr.2018.07.034
Citation Information
Pranav Joshi, Kyeong Nam Yu, Soo Yeon Kang, Seok Joon Kwon, et al.. "3D-Cultured Neural Stem Cell Microarrays on a Micropillar Chip for High-Throughput Developmental Neurotoxicology" Experimental Cell Research Vol. 370 Iss. 2 (2018) p. 680 - 691 ISSN: 00144827
Available at: http://works.bepress.com/Chandrasekhar-Kothapalli/7/