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Mol. Cells 2013; 35(5): 456-461

Published online May 16, 2013

https://doi.org/10.1007/s10059-013-0083-0

© The Korean Society for Molecular and Cellular Biology

The Establishment of Mouse Embryonic Stem Cell Cultures on 96-Well Plates for High-Throughput Screening

Meeyoung Cho, Tae-Jun Cho, Jeong Mook Lim, Gene Lee, and Jaejin Cho

1Department of Dental Regenerative Biotechnology, 2Dental Research Institute, 3World Class University, Department of Agricultural Biotechnology, 4Laboratory of Molecular Genetics, School of Dentistry, Seoul National University, Seoul 110-749, Korea

Received: March 11, 2013; Revised: March 28, 2013; Accepted: March 29, 2013

Abstract

Embryonic stem (ES) cells can be valuable for monitoring differentiation processes and for improving applications in basic developmental biology. The application of ES cells can be a useful tool for drug discovery and toxi-cology. Therefore, we suggest the high-throughput screening (HTS) system based on ES cells in this study. Firstly, we optimized the feeder-free condition and seeding cell number which can maintained for at least 7 days without over-con-fluency. We analyzed the system by cell viability, proliferation activity, RT-PCR and morphologic/immunohistoche-mical evaluations. The optimal cell seeding number was 30/well that was maintained the typical colonial morphology over 9 d with 1,000 U/ml LIF in the limited space. The cell in optimized condition expressed ALP, SSEA-1, Oct 4 and Nanog and the genetic expressions showed similar to protein expressions. The cell lineage marker expressions showed faint or none. The cell viability and proliferation activity were increased in time-dependent manner in our optimized HTS system. In conclusion, the novel HTS system using ES cells can by useful for developing mod-els for drug discovery as well as toxicological screening in the near future.

Keywords 96-well plate, embryonic stem cell, feeder-free, high-throughput screening

Article

Research Article

Mol. Cells 2013; 35(5): 456-461

Published online May 31, 2013 https://doi.org/10.1007/s10059-013-0083-0

Copyright © The Korean Society for Molecular and Cellular Biology.

The Establishment of Mouse Embryonic Stem Cell Cultures on 96-Well Plates for High-Throughput Screening

Meeyoung Cho, Tae-Jun Cho, Jeong Mook Lim, Gene Lee, and Jaejin Cho

1Department of Dental Regenerative Biotechnology, 2Dental Research Institute, 3World Class University, Department of Agricultural Biotechnology, 4Laboratory of Molecular Genetics, School of Dentistry, Seoul National University, Seoul 110-749, Korea

Received: March 11, 2013; Revised: March 28, 2013; Accepted: March 29, 2013

Abstract

Embryonic stem (ES) cells can be valuable for monitoring differentiation processes and for improving applications in basic developmental biology. The application of ES cells can be a useful tool for drug discovery and toxi-cology. Therefore, we suggest the high-throughput screening (HTS) system based on ES cells in this study. Firstly, we optimized the feeder-free condition and seeding cell number which can maintained for at least 7 days without over-con-fluency. We analyzed the system by cell viability, proliferation activity, RT-PCR and morphologic/immunohistoche-mical evaluations. The optimal cell seeding number was 30/well that was maintained the typical colonial morphology over 9 d with 1,000 U/ml LIF in the limited space. The cell in optimized condition expressed ALP, SSEA-1, Oct 4 and Nanog and the genetic expressions showed similar to protein expressions. The cell lineage marker expressions showed faint or none. The cell viability and proliferation activity were increased in time-dependent manner in our optimized HTS system. In conclusion, the novel HTS system using ES cells can by useful for developing mod-els for drug discovery as well as toxicological screening in the near future.

Keywords: 96-well plate, embryonic stem cell, feeder-free, high-throughput screening

Mol. Cells
Jun 30, 2023 Vol.46 No.6, pp. 329~398
COVER PICTURE
The cellular proteostasis network is adaptively modulated upon cellular stress, thereby protecting cells from proteostasis collapse. Heat shock induces the translocation of misfolded proteins and the chaperone protein HSP70 into nucleolus, where nuclear protein quality control primarily occurs. Nuclear RNA export factor 1 (green), nucleolar protein fibrillarin (red), and nuclei (blue) were visualized in NIH3T3 cells under basal (left) and heat shock (right) conditions (Park et al., pp. 374-386).

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