Journal of Animal Reproduction and Biotechnology (한국동물생명공학회지)

The Korean Society of Animal Reproduction and Biotechnology (한국동물생명공학회)
권호 표지
DOI QR코드

DOI QR Code

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)
  • Received : 2019.12.05
  • Accepted : 2020.02.06
  • Published : 2020.03.31
Download PDF
⟨ Previous Next ⟩

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

References

  1. Amano M, Nakayama M, Kaibuchi K. 2010. Rho-kinase/ROCK: a key regulator of the cytoskeleton and cell polarity. Cytoskeleton. 67:545-554. https://doi.org/10.1002/cm.20472
  2. Cavallini A, Micieli G, Marcheselli S, Quaglini S. 2003. Role of monitoring in management of acute ischemic stroke patients. Stroke. 34:2599-2603. https://doi.org/10.1161/01.str.0000094423.34841.bb
  3. 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. https://doi.org/10.1517/13543784.2013.778242
  4. 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. https://doi.org/10.1159/000430118
  5. 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. https://doi.org/10.1016/j.ijcard.2015.08.118
  6. 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. https://doi.org/10.1038/35070009
  7. Gao S, McGarry M, Latham KE, Wilmut I. 2003. Cloning of mice by nuclear transfer. Cloning & Stem Cells. 5:287-294. https://doi.org/10.1089/153623003772032790
  8. 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. https://doi.org/10.1016/S0015-0282(00)00518-5
  9. 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.
  10. 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. https://doi.org/10.2174/1874467209666160112125446
  11. 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. https://doi.org/10.1095/biolreprod.108.070102
  12. 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. https://doi.org/10.1038/nature13134
  13. 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. https://doi.org/10.1002/1526-968X(200011/12)28:3/4<156::AID-GENE100>3.0.CO;2-T
  14. Leyva V, Buckrell B, Walton J. 1998. Follicular activity and ovulation regulated by exogenous progestagen and PMSG in anestrous ewes. Theriogenology. 50:377-393. https://doi.org/10.1016/S0093-691X(98)00147-2
  15. 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.
  16. 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. https://doi.org/10.1007/s00228-011-1100-x
  17. 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. https://doi.org/10.1038/nature13551
  18. 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. https://doi.org/10.12750/JET.2017.32.3.81
  19. 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. https://doi.org/10.1093/oxfordjournals.humrep.a138171
  20. 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. https://doi.org/10.1039/c1ob05332a
  21. Polejaeva I, Campbell K. 2000. New advances in somatic cell nuclear transfer: application in transgenesis. Theriogenology. 53:117-126. https://doi.org/10.1016/S0093-691X(99)00245-9
  22. Roh S. 2012. Somatic cell nuclear transfer in rodents, the little big animals. Journal of Animal Reproduciton and Biotechnology. 27:205-209.
  23. 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.
  24. 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. https://doi.org/10.1016/j.cell.2013.05.006
  25. Tsung H, Mummery CL. 1990. Effects of feeder layer and BRL conditioned medium on mouse embryonic stem cells. Cell Research. 1:35. https://doi.org/10.1038/cr.1990.5
  26. 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. https://doi.org/10.3727/096504003108748500
  27. 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. https://doi.org/10.1038/nbt1310
  28. 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. https://doi.org/10.1038/nature06968
  29. 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. https://doi.org/10.1038/sj.cr.7310139