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http://dx.doi.org/10.12750/JARB.35.1.21

Fasudil Increases the Establishment of Somatic Cell Nuclear Transfer Embryonic Stem Cells in Mouse  

So, Seongjun (Department of Convergence Medicine & Stem Cell Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine)
Karagozlu, Mustafa Zafer (Department of Convergence Medicine & Stem Cell Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine)
Lee, Yeonmi (Department of Convergence Medicine & Stem Cell Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine)
Kang, Eunju (Department of Convergence Medicine & Stem Cell Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine)
Publication Information
Journal of Animal Reproduction and Biotechnology / v.35, no.1, 2020 , pp. 21-27 More about this Journal
Abstract
Somatic cell nuclear transfer derived embryonic stem cells (NT-ESCs) have significant advantages in various fields such as genetics, embryology, stem cell science, and regenerative medicine. However, the poor establishment of NT-ESCs hinders various research. Here, we applied fasudil, a Rho-associated kinase (ROCK) inhibitor, to develop somatic cell nuclear transfer (SCNT) embryos and establish NT-ESCs. In the study, MII oocytes were isolated from female B6D2F1 mice and performed SCNT with mouse embryonic fibroblasts (MEFs). The reconstructed NT-oocytes were activated artificially, and cultured to blastocysts in KSOM supplemented with 10 μM fasudil. Further, the blastocysts were seeded on inactivated MEFs in embryonic stem cell medium supplemented with 10 μM fasudil. A total of 26% of embryos formed into blastocysts in the fasudil treated group, while this ratio was 44% in the fasudil free control group. On the other hand, 30% of blastocysts were established NT-ESCs after exposure of fasudil, which was significantly higher than the control group (10%). The results suggest that fasudil reduced blastocyst development after SCNT due to inhibition of 2 cell cleavage while improved the establishment of NT-ESCs through the anti-apoptotic pathway.
Keywords
embryo development; embryonic stem cells; fasudil; somatic cells nuclear transfer;
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1 Wakayama T. 2003. Cloned mice and embryonic stem cell lines generated from adult somatic cells by nuclear transfer. Oncology Research Featuring Preclinical and Clinical Cancer Therapeutics. 13:309-314.   DOI
2 Watanabe K, Ueno M, Kamiya D, Nishiyama A, Matsumura M, Wataya T, Takahashi JB, Nishikawa S, Nishikawa S-i, Muguruma K. 2007. A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nature Biotechnology. 25:681.   DOI
3 Ying QL, Wray J, Nichols J, Batlle-Morera L, Doble B, Woodgett J, Cohen P, Smith A. 2008. The ground state of embryonic stem cell self-renewal. Nature. 453:519-523.   DOI
4 Zhao C, Yao R, Hao J, Ding C, Fan Y, Dai X, Li W, Hai T, Liu Z, Yu Y. 2007. Establishment of customized mouse stem cell lines by sequential nuclear transfer. Cell Research. 17:80.   DOI
5 Amano M, Nakayama M, Kaibuchi K. 2010. Rho-kinase/ROCK: a key regulator of the cytoskeleton and cell polarity. Cytoskeleton. 67:545-554.   DOI
6 Cavallini A, Micieli G, Marcheselli S, Quaglini S. 2003. Role of monitoring in management of acute ischemic stroke patients. Stroke. 34:2599-2603.   DOI
7 Chen M, Liu A, Ouyang Y, Huang Y, Chao X, Pi R. 2013. Fasudil and its analogs: a new powerful weapon in the long war against central nervous system disorders? Expert Opinion on Investigational Drugs. 22:537-550.   DOI
8 Coleman ML, Sahai EA, Yeo M, Bosch M, Dewar A, Olson MF. 2001. Membrane blebbing during apoptosis results from caspase-mediated activation of ROCK I. Nature Cell Biology. 3:339.   DOI
9 Chen S, Luo M, Zhao Y, Zhang Y, He M, Cai W, Liu A. 2015. Fasudil stimulates neurite outgrowth and promotes differentiation in C17. 2 neural stem cells by modulating notch signalling but not autophagy. Cellular Physiology and Biochemistry. 36:531-541.   DOI
10 Cheng Y-T, Yeih D-F, Liang S-M, Chien C-Y, Yu Y-L, Ko B-S, Jan Y-J, Kuo C-C, Sung L-Y, Shyue S-K. 2015. Rho-associated kinase inhibitors promote the cardiac differentiation of embryonic and induced pluripotent stem cells. International Journal of Cardiology. 201:441-448.   DOI
11 Gao S, McGarry M, Latham KE, Wilmut I. 2003. Cloning of mice by nuclear transfer. Cloning & Stem Cells. 5:287-294.   DOI
12 Gardner DK, Lane M, Stevens J, Schlenker T, Schoolcraft WB. 2000. Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertility and Sterility. 73:1155-1158.   DOI
13 Kang E, Wu G, Ma H, Li Y, Tippner-Hedges R, Tachibana M, Sparman M, Wolf DP, Scholer HR, Mitalipov S. 2014. Nuclear reprogramming by interphase cytoplasm of two-cell mouse embryos. Nature. 509:101-104.   DOI
14 Hu Y, Li X, Huang G, Wang J, Lu W. 2019. Fasudil may induce the differentiation of bone marrow mesenchymal stem cells into neuron-like cells via the Wnt/${\beta}$-catenin pathway. Molecular Medicine Reports. 19:3095-3104.
15 Jiao X, Ashtari N, Rahimi-Balaei M, Min Chen Q, Badbezanchi I, Shojaei S, Marzban A, Mirzaei N, Chung S, Guan T. 2017. Mevalonate cascade and neurodevelopmental and neurodegenerative diseases: Future targets for therapeutic application. Current Molecular Pharmacology. 10:115-140.   DOI
16 Jin X, Chandrakanthan V, Morgan H, O'neill C. 2009. Preimplantation embryo development in the mouse requires the latency of TRP53 expression, which is induced by a ligandactivated PI3 kinase/AKT/MDM2-mediated signaling pathway. Biology of Reproduction. 80:286-294.   DOI
17 Kawase E, Yamazaki Y, Yagi T, Yanagimachi R, Pedersen RA. 2000. Mouse embryonic stem (ES) cell lines established from neuronal cell-derived cloned blastocysts. Genesis. 28:156-163.   DOI
18 Leyva V, Buckrell B, Walton J. 1998. Follicular activity and ovulation regulated by exogenous progestagen and PMSG in anestrous ewes. Theriogenology. 50:377-393.   DOI
19 Liu A-j, Ling F, Wang D, Wang Q, LU X-d, Liu Y-l. 2011. Fasudil inhibits platelet-derived growth factor-induced human pulmonary artery smooth muscle cell proliferation by upregulation of p27kip1 via the ERK signal pathway. Chinese Medical Journal. 124:3098-3104.
20 Liu GJ, Wang ZJ, Wang YF, Xu LL, Wang XL, Liu Y, Luo GJ, He GH, Zeng YJ. 2012. Systematic assessment and meta-analysis of the efficacy and safety of fasudil in the treatment of cerebral vasospasm in patients with subarachnoid hemorrhage. European Journal of Clinical Pharmacology. 68:131-139.   DOI
21 Palecek J, Zweigerdt R, Olmer R, Martin U, Kirschning A, Drager G. 2011. A practical synthesis of Rho-Kinase inhibitor Y-27632 and fluoro derivatives and their evaluation in human pluripotent stem cells. Organic & Biomolecular Chemistry. 9:5503-5510.   DOI
22 Ma H, Morey R, O'Neil RC, He Y, Daughtry B, Schultz MD, Hariharan M, Nery JR, Castanon R, Sabatini K. 2014. Abnormalities in human pluripotent cells due to reprogramming mechanisms. Nature. 511:177.   DOI
23 Moon J, Jung M, Roh S. 2017. Comparison of developmental efficiency of murine somatic cell nuclear transfer protocol. Journal of Animal Reproduciton and Biotechnology. 32:81-86.   DOI
24 Neev J, Gonzalez A, Licciardi F, Alikani M, Tadir Y, Berns M, Cohen J. 1993. Opening of the mouse zona pellucida by laser without a micromanipulator. Human Reproduction. 8:939-944.   DOI
25 Polejaeva I, Campbell K. 2000. New advances in somatic cell nuclear transfer: application in transgenesis. Theriogenology. 53:117-126.   DOI
26 Roh S. 2012. Somatic cell nuclear transfer in rodents, the little big animals. Journal of Animal Reproduciton and Biotechnology. 27:205-209.
27 Tsung H, Mummery CL. 1990. Effects of feeder layer and BRL conditioned medium on mouse embryonic stem cells. Cell Research. 1:35.   DOI
28 So S, Lee Y, Park J, Lee JY, Kim D, Hwang J, Shin J, Choi J, Han Y, Kang S, Dutton JR, Seo EJ, Lee BH, Kim CJ, Mitalipov S, Oh S, Kang E. 2019. The Rho-associated kinase inhibitor fasudil can replace Y-27632 for use in human pluripotent stem cell research. Preprints. doi: 10.20944.
29 Tachibana M, Amato P, Sparman M, Gutierrez NM, Tippner-Hedges R, Ma H, Kang E, Fulati A, Lee H-S, Sritanaudomchai H. 2013. Human embryonic stem cells derived by somatic cell nuclear transfer. Cell 153:1228-1238.   DOI