Efficient production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation

Abstract

A large number of hDAF transgenic pigs to be used for xenotransplantation research were generated by using sperm-mediated gene transfer (SMGT). The efficiency of transgenesis obtained with SMGT was much greater than with any other method. In the experiments reported, up to 80% of pigs had the transgene integrated into the genome. Most of the pigs carrying the hDAF gene transcribed it in a stable manner (64%). The great majority of pigs that transcribed the gene expressed the protein (83%). The hDAF gene was transmitted to progeny. Expression was stable and found in caveolae as it is in human cells. The expressed gene was functional based on in vitro experiments performed on peripheral blood mononuclear cells. These results show that our SMGT approach to transgenesis provides an efficient procedure for studies involving large animal models.SMGT ͉ hyperacute rejection ͉ swine

Evaluation of swine fertilisation medium (SFM) efficiency in preserving spermatozoa quality during long-term storage in comparison to four commercial swine extenders

“…For this reason, there has been increased interest in developing new extenders and in establishing the optimal storage conditions for diluted spermatozoa. In the last few decades, we utilised a homemade diluent (swine fertilisation medium (SFM)) for swine spermatozoa manipulation and biotechnological applications as the production of transgenic pigs utilising the sperm-mediated gene transfer technique (Lavitrano et al, 2002;Lavitrano et al, 2003), with good fertility results as well as in AI or surgical or laparoscopic inseminations Manzini et al, 2006). Nevertheless, this SFM has never been evaluated methodically in comparison to other well-defined commercial diluents.…”

Status of transgenic chicken models for developmental biology

“…The sperm that successfully fertilizes an egg carries the transgene of interest into the egg, the transgene becomes incorporated into the resulting embryo and hatched chicks. Recently, spermmediated transfer has been reported to be a reproducible way to make transgenic mammals (Lavitrano et al, 2002(Lavitrano et al, , 2003. However, sperm-mediated gene transfer has not yet provided any convincing data to suggest that a transgene of interest can successfully be incorporated into the avian germline.…”

Relations between animal transgenesis and reproduction

“…This extremely simple technique gave birth to transgenic animals of different species. The method which originally appeared poorly reproducible has been greatly improved by eliminating the DNAse which is abundant in seminal plasma and on the sperm surface [10].…”