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High-content imaging of 3D-cultured neural stem cells on a 384-pillar plate for the assessment of cytotoxicity.
Toxicol In Vitro 2020; 65:104765TV

Abstract

The assessment of neurotoxicity has been performed traditionally with animals. However, in vivo studies are highly expensive and time-consuming, and often do not correlate to human outcomes. Thus, there is a need for cost-effective, high-throughput, highly predictive alternative in vitro test methods based on early markers of mechanisms of toxicity. High-content imaging (HCI) assays performed on three-dimensionally (3D) cultured cells could provide better understanding of the mechanism of toxicity needed to predict neurotoxicity in humans. However, current 3D cell culture systems lack the throughput required for screening neurotoxicity against a large number of chemicals. Therefore, we have developed miniature 3D neural stem cell (NSC) culture on a unique 384-pillar plate, which is complementary to conventional 384-well plates. Mitochondrial membrane impairment, intracellular glutathione level, cell membrane integrity, DNA damage, and apoptosis have been tested against 3D-cultured ReNcell VM on the 384-pillar plate with four model compounds rotenone, 4-aminopyridine, digoxin, and topotecan. The HCI assays performed in 3D-cultured ReNcell VM on the 384-pillar plates were highly robust and reproducible as indicated by the average Z' factor of 0.6 and CV values around 12%. From concentration-response curves and IC50 values, mitochondrial membrane impairment appears to be the early stage marker of cell death by the compounds.

Authors+Show Affiliations

Department of Chemical and Biomedical Engineering, Cleveland State University, 1960 East 24th Street, Cleveland, OH 44115, United States of America.Department of Chemical and Biomedical Engineering, Cleveland State University, 1960 East 24th Street, Cleveland, OH 44115, United States of America.Department of Chemical and Biomedical Engineering, Cleveland State University, 1960 East 24th Street, Cleveland, OH 44115, United States of America.Department of Chemical and Biomedical Engineering, Cleveland State University, 1960 East 24th Street, Cleveland, OH 44115, United States of America.Department of Chemical and Biomedical Engineering, Cleveland State University, 1960 East 24th Street, Cleveland, OH 44115, United States of America. Electronic address: m.lee68@csuohio.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31923580

Citation

Joshi, Pranav, et al. "High-content Imaging of 3D-cultured Neural Stem Cells On a 384-pillar Plate for the Assessment of Cytotoxicity." Toxicology in Vitro : an International Journal Published in Association With BIBRA, vol. 65, 2020, p. 104765.
Joshi P, Kang SY, Yu KN, et al. High-content imaging of 3D-cultured neural stem cells on a 384-pillar plate for the assessment of cytotoxicity. Toxicol In Vitro. 2020;65:104765.
Joshi, P., Kang, S. Y., Yu, K. N., Kothapalli, C., & Lee, M. Y. (2020). High-content imaging of 3D-cultured neural stem cells on a 384-pillar plate for the assessment of cytotoxicity. Toxicology in Vitro : an International Journal Published in Association With BIBRA, 65, p. 104765. doi:10.1016/j.tiv.2020.104765.
Joshi P, et al. High-content Imaging of 3D-cultured Neural Stem Cells On a 384-pillar Plate for the Assessment of Cytotoxicity. Toxicol In Vitro. 2020 Jan 7;65:104765. PubMed PMID: 31923580.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High-content imaging of 3D-cultured neural stem cells on a 384-pillar plate for the assessment of cytotoxicity. AU - Joshi,Pranav, AU - Kang,Soo-Yeon, AU - Yu,Kyeong-Nam, AU - Kothapalli,Chandrasekhar, AU - Lee,Moo-Yeal, Y1 - 2020/01/07/ PY - 2019/07/19/received PY - 2019/12/20/revised PY - 2020/01/05/accepted PY - 2020/1/11/pubmed PY - 2020/1/11/medline PY - 2020/1/11/entrez KW - 384-Pillar plate KW - High-content imaging (HCI) KW - High-throughput screening (HTS) KW - Neural stem cell (NSC) KW - Neurotoxicity KW - Three-dimensional (3D) cell culture SP - 104765 EP - 104765 JF - Toxicology in vitro : an international journal published in association with BIBRA JO - Toxicol In Vitro VL - 65 N2 - The assessment of neurotoxicity has been performed traditionally with animals. However, in vivo studies are highly expensive and time-consuming, and often do not correlate to human outcomes. Thus, there is a need for cost-effective, high-throughput, highly predictive alternative in vitro test methods based on early markers of mechanisms of toxicity. High-content imaging (HCI) assays performed on three-dimensionally (3D) cultured cells could provide better understanding of the mechanism of toxicity needed to predict neurotoxicity in humans. However, current 3D cell culture systems lack the throughput required for screening neurotoxicity against a large number of chemicals. Therefore, we have developed miniature 3D neural stem cell (NSC) culture on a unique 384-pillar plate, which is complementary to conventional 384-well plates. Mitochondrial membrane impairment, intracellular glutathione level, cell membrane integrity, DNA damage, and apoptosis have been tested against 3D-cultured ReNcell VM on the 384-pillar plate with four model compounds rotenone, 4-aminopyridine, digoxin, and topotecan. The HCI assays performed in 3D-cultured ReNcell VM on the 384-pillar plates were highly robust and reproducible as indicated by the average Z' factor of 0.6 and CV values around 12%. From concentration-response curves and IC50 values, mitochondrial membrane impairment appears to be the early stage marker of cell death by the compounds. SN - 1879-3177 UR - https://www.unboundmedicine.com/medline/citation/31923580/High-content_imaging_of_3D-cultured_neural_stem_cells_on_a_384-pillar_plate_for_the_assessment_of_cytotoxicity L2 - https://linkinghub.elsevier.com/retrieve/pii/S0887-2333(19)30558-2 DB - PRIME DP - Unbound Medicine ER -