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KMID : 0613820190290080895
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Journal of Life Science
2019 Volume.29 No. 8 p.895 ~ p.903
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Improvement of Cell Viability Using a Rho-associated Protein Kinase (ROCK) Inhibitor in Human Dental Papilla derived Single-induced Pluripotent Stem Cells
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Shim Yoo-Jin
Kang Young-Hoon
Kim Hyeon-Ji
Kim Mi-Jeong
Lee Hyeon-Jeong
Son Young-Bum
Lee Sung-Ho
Jeon Byeong-Gyun
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Abstract
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The aim of the present study was to improve the cell viability of human dental papilla derived single-induced pluripotent stem cells (iPSCs) using a Rho-associated protein kinase (ROCK) inhibitor, Y-27632. The iPSCs were produced using an episomal plasmid-based reprogramming method. After cell separation using trypsin, the iPSCs were treated with 0, 0.5, 1, 2.5, 5, 7.5, or 10 μM Y-27632 for 5 d. Cell viability increased significantly following the 5 μM Y-27632 treatment (p<0.05). When the iPSCs were exposed to medium containing 10 μM Y-27632 for 0, 1, 2, 3, 4, and 5 d, the cell viability rate increased significantly in accordance with the cell viability rate (p<0.05). To evaluate the effect of the Y-27632 treatment on stemness characteristics, the expression of stem cell-specific transcripts and telomerase activity were investigated in the iPSCs treated with 10 μM Y-27632 for 5 d. The expression levels of stem cell-specific transcripts, such as OCT-4, NONOG, and SOX-2, and telomerase activity were not significantly different in the iPSCs treated with 10 μM Y-27632 as compared with those of untreated control iPSCs (p>0.05). Taken together, the results demonstrated that cell viability can be improved by treatment with the ROCK inhibitor Y-27632, without losing iPSC stemness characteristics.
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KEYWORD
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Anoikis, human, iPSCs, stem cells, Y-27632
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