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
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 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.
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
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
Yujeong Oh, and Jiwon Jang
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