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First published online February 2, 2006
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2005-0391v1
24/6/1450    most recent
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Submitted on August 13, 2005
Accepted on January 23, 2006

Embryonic Stem Cells

Improved Safety of Hematopoietic Transplantation with Monkey ES Cells in the Allogeneic Setting

Hiroaki Shibata 1, Naohide Ageyama 1, Yujiro Tanaka 2, Yukiko Kishi 2, Kyoko Sasaki 2, Shinichiro Nakamura 1, Shin-ichi Muramatsu 3, Satoshi Hayashi 4, Yoshihiro Kitano 5, Keiji Terao 1, Yutaka Hanazono 2*

1 Tsukuba Primate Research Center, National Institute of Biomedical Innovation, Ibaraki, Japan
2 Center for Molecular Medicine, Jichi Medical School, Tochigi, Japan
3 Department of Neurology, Jichi Medical University, Tochigi, Japan
4 Department of Obstetrics and Gynecology, National Center for Child Hearth and Development, Tokyo, Japan
5 Department of Surgery, National Center for Child Hearth and Development, Tokyo, Japan

* To whom correspondence should be addressed. E-mail: hanazono{at}jichi.ac.jp.


  Abstract

Cynomolgus monkey embryonic stem cell (cyESC)-derived in vivo hematopoiesis was examined in an allogeneic transplantation model. cyESCs were induced to differentiate into the putative hematopoietic precursors in vitro and the cells were transplanted into the fetal cynomolgus liver around the end of first trimester (n = 3). Although cyESC-derived hematopoietic colony-forming cells were detected in the newborns (4.1 and 4.7%), a teratoma developed in all newborns. The risk of tumor formation was high in this allogeneic transplantation model, given that tumors were hardly observed in immunodeficient mice or fetal sheep that had been xeno-transplanted with the same cyESC-derivatives. It turned out that the cyESC-derived donor cells included a residual undifferentiated fraction positive for stage-specific embryonic antigen (SSEA)-4 (38.2 ± 10.3%) despite the rigorous differentiation culture. When an SSEA-4-negative fraction was transplanted (n = 6), the teratoma was no longer observed while the cyESC-derived hematopoietic engraftment was unperturbed (2.3-5.0%). SSEA-4 is therefore a clinically-relevant pluripotency marker of primate embryonic stem (ES) cells. Purging pluripotent cells with this surface marker would be a promising method of producing clinical progenitor cell preparations using human ES cells.

Key Words. Non-human primate embryonic stem cell, Primate allogeneic transplantation, In utero transplantation, Hematopoiesis, Teratoma, Tumor prevention




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