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Mol. Hum. Reprod. Advance Access originally published online on October 21, 2008
Molecular Human Reproduction 2008 14(11):627-634; doi:10.1093/molehr/gan061
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© The Author 2008. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org
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Rho-associated kinase inhibitor Y-27632 promotes survival of cynomolgus monkey embryonic stem cells

Toshiyuki Takehara1, Takeshi Teramura2,3,*, Yuta Onodera2, Ryo Kakegawa1, Naoto Fukunaga1, Makoto Takenoshita4, Norimasa Sagawa3, Kanji Fukuda2,5 and Yoshihiko Hosoi1

1Graduate School of Biology-Oriented Science and Technology, Kinki University, Nishimitani, Kinokawa, Wakayama 649-6493, Japan 2Institute of Advanced Clinical Medicine, Kinki University School of Medicine, Ohno-Higashi, Osaka-Sayama, Osaka 589-8511, Japan 3Department of Obstetrics and Gynecology, Graduate School of Medicine, Mie University, Edobashi 2-174, Tsu, Mie 514-8507, Japan 4Wakayama Research Institute, Keari Co., Ltd, . Yamauchi 514, Sumida, Hashimoto, Wakayama 648-0003, Japan 5Department of Orthopaedic Surgery, Kinki University School of Medicine, Ohno-Higashi, Osaka-Sayama, Osaka 589-8511, Japan

6 Correspondence address. Institute of Advanced Clinical Medicine, Kinki University School of Medicine, Ohno-Higashi, Osaka-Sayama, Osaka 589-8511, Japan. Tel: +81-72-36-0221; Fax: +81-72-366-0206; E-mail: teramura{at}med.kindai.ac.jp

Non-human primates are suitable models for preclinical research aimed at cell-replacement therapies. Recently, it has been reported that Rho-associated kinase inhibitor Y-27632 markedly reduced dissociation-induced apoptosis of human embryonic stem (hES) cells, and is expected as a novel supplement for hES cell maintenance or differentiation inductions; however, the effects of the chemical are still to be determined in model animals. Here, we demonstrated the effect of Y-27632 on cynomolgus monkey ES (cyES) cells. Also, in cyES cells, Y-27632 treatment dramatically improved the efficiency of colony formation from single cells without affecting the pluripotent state and karyotype. Y-27632 supplementation was also effective for feeder-free culture and differentiation induction. Neural stem cells directly induced from cyES cells could give rise to neurons, astrocytes and dopamine producing cells. The present result not only suggests that the chemical was effective for improving the culture system of primate ES cells, but also the similarity between cyES and hES cells regarding the reactions to the chemical, which might be further evidence that cyES cells are superior models for hES cells.

Key words: primate/embryonic stem cells/pluripotency/differentiation

Submitted on February 20, 2008; resubmitted on October 14, 2008; accepted on October 16, 2008.


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