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Cytolytic assessment of hyperacute rejection and production of nuclear transfer embryos using hCD46-transgenic porcine embryonic germ cells

Published online by Cambridge University Press:  01 May 2009

Ji Young Won
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
Kwang Sung Ahn
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
Alice M. Sorrell
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea. Metabolic Research Laboratories, Institute of Metabolic Science, University of Cambridge, Addenbrooke's Treatment Centre, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 0QQ, UK.
Susa Shin
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
Soon Young Heo
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
Jee Hyun Kang
Affiliation:
Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
Jin-Ki Park
Affiliation:
Animal Biotechnology Division, National Institute of Animal Science, Suwon, Korea.
Won-Kyong Chang
Affiliation:
Animal Biotechnology Division, National Institute of Animal Science, Suwon, Korea.
Hosup Shim*
Affiliation:
Department of Physiology, Dankook University School of Medicine, San 29 Anseo-dong, Cheonan, Chungnam 330–714, South Korea. Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
*
All correspondence to: Hosup Shim. Department of Physiology, Dankook University School of Medicine, San 29 Anseo-dong, Cheonan, Chungnam 330–714, South Korea. Tel: +82 41 550 3865. Fax: +82 41 565 6167. e-mail: shim@dku.edu

Summary

Human complement regulatory protein hCD46 may reduce the hyperacute rejection (HAR) in pig-to-human xenotransplantation. In this study, an hCD46 gene was introduced into porcine embryonic germ (EG) cells. Treatment of human serum did not affect the survival of hCD46-transgenic EG cells, whereas the treatment significantly reduced the survival of non-transgenic EG cells (p < 0.01). The transgenic EG cells presumably capable of alleviating HAR were transferred into enucleated oocytes. Among 235 reconstituted oocytes, 35 (14.9%) developed to the blastocyst stage. Analysis of individual embryos indicated that 80.0% (28/35) of embryos contained the transgene hCD46. The result of the present study demonstrates resistance of hCD46-transgenic EG cells against HAR, and the usefulness of the transgenic approach may be predicted by this cytolytic assessment prior to actual production of transgenic pigs. Subsequently performed EG cell nuclear transfer gave rise to hCD46-transgenic embryos. Further study on the transfer of these embryos to recipients may produce hCD46-transgenic pigs.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2008

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