title: Cytolytic assessment of hyperacute rejection and production of nuclear transfer embryos using hCD46-transgenic porcine embryonic germ cells.
abstract: 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.
authors.affiliation: Department of Physiology, Dankook University School of Medicine, Cheonan, Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Won
authors.forename: Kwang Sung
authors.initials: KS
authors.lastname: Ahn
authors.forename: Alice M
authors.initials: AM
authors.lastname: Sorrell
authors.forename: Susa
authors.initials: S
authors.lastname: Shin
authors.forename: Soon Young
authors.initials: SY
authors.lastname: Heo
authors.forename: Jee Hyun
authors.initials: JH
authors.lastname: Kang
authors.forename: Jin-Ki
authors.initials: JK
authors.lastname: Park
authors.forename: Won-Kyong
authors.initials: WK
authors.lastname: Chang
authors.forename: Hosup
authors.initials: H
authors.lastname: Shim
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000502:physiology*; D047108:Embryonic Development; D018014:Gene Transfer Techniques; D006801:Humans; D051916:Membrane Cofactor Protein / Q000235:genetics* / Q000378:metabolism; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000502:physiology*; D013552:Swine / Q000235:genetics*; D019076:Transgenes
chemical_list: C497857:CD46 protein, human; D051916:Membrane Cofactor Protein