Angiogenesis is the process that drives blood vessel development in growing tissues in response to the local production of angiogenic factors. With t

A large number ofhDAFtransgenic 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 thehDAFgene transcribed it in a stable manner (64%). The great majority of pigs that transcribed the gene expressed the protein (83%). ThehDAFgene was transmitted to progeny. Expression was stable and found incaveolaeas it is in human cells. The expressed gene was functional based onin vitroexperiments 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.

The similarities between swine and humans in physiological and genomic patterns, and the great correlation in size and anatomy, make pigs extremely useful in preclinical studies. New-born piglets can represent a model for congenital and genetic diseases in new-born children. It is known that piglets may have significant differences in clinicopathological results compared to adult pigs. Therefore, adult laboratory reference intervals cannot be applied to piglets. The aim of this study was to compare haematological and chemical variables in piglets of two ages and determinate age-related reference intervals for commercial hybrid young pigs. Blood samples were collected under general anaesthesia from 130 animals divided into five- (P5) and 30- (P30) day-old piglets. Only P30 animals were treated with parenteral iron after birth. Samples were analysed using automated haematology (ADVIA 2120) and chemistry analysers, and age-related reference intervals were calculated. Significant higher values of RBC, Hb and HCT were observed in P30 animals when compared to P5, with an opposite trend for MCV. These results were associated with a reduction of the RBC regeneration process and the thrombopoietic response. The TSAT and TIBC were significantly higher in P30 compared to P5; however, piglets remained iron deficient compared to adult reference intervals reported previously. In conclusion, this paper emphasises the high variability occurring in clinicopathological variables between new-born and 30-day-old pigs, and between piglets and adult pigs. This study provides valuable reference data for piglets at precise ages and could be used in the future as historical control improving the Reduction in animal experiments, as suggested by the 3Rs principle.

Ejaculated sperm cells from both bovine and swine animals depleted of the seminal fluid were used as vectors for transferring different plasmid DNAs into eggs during the fertilization process. PCR screening of about 2000 blastocysts showed that transformed embryos were obtained in both animal species with varying efficiencies, and that the rate of transformation depended on the plasmid used. The most efficient rate was obtained in both species with the pSV2CAT construct, yielding 22% positive blastocysts in bovine and 5.7% in swine respectively. Artificial insemination of ten sows with boar sperm cells that had been preincubated with pSV2CAT DNA produced 82 offspring. Southern blot analysis of the DNAs extracted from the animal tails showed that five animals were transgenic and contained sequences complementary to pSV2CAT DNA that appeared to be rearranged compared to the original plasmid. These results indicate that sperm‐mediated DNA transfer protocols can be successfully adapted for the generation of genetically transformed livestock.