22 MALFORMATIONS FOUND BY AUTOPSY OF CLONED AND TRANSGENIC PIGLETS OF DIFFERENT BREEDS

Viability of cloned and transgenic piglets is seriously compromised and one obvious reason could be malformations. The aim of the present study was therefore to describe gross pathological conditions in dead pre-weaned piglets born after transfer to Large White (LW) recipients of cloned (LW donor cells) or transgenic (Yucatan or Gottingen donor cells) embryos. Donor cells were fibroblasts and the Gottingen and Yucatan cells were made transgenic with 1 of 5 genes known to dispose for different human diseases. Handmade cloning was used to produce embryos that after 5 to 6 days of in vitro culture were transferred surgically to 108 LW sows 4 days after their natural heat. Of these, 21 sows delivered cloned LW piglets, whereas 17 and 16 sows, respectively, delivered transgenic Gottingen and Yucatan piglets. Stillborn and dead pre-weaned piglets were necropsied and malformations registered. Data were analysed by Fisher's exact test with a significance level of P < 0.05. In the 54 litters, total litter size ranged from 1 to 22 piglets (mean 5.4 ± 0.5) and the overall mortality rate until weaning on day 28 was 59%. Malformations were found in piglets from 38 litters where an average of 35% of the piglets showed malformations (between 8 and 100%). In those litters, 1 to 7 piglets had 1, 2, or several malformations (Table 1). The malformation rate in the autopsied transgenic Gottingen was 58% and in Yucatan 46%; these were significantly higher than in the autopsied cloned LW piglets with 18%. Some of the malformations seemed to be related to breed and/or transgene; for instance, heart malformations were most frequent in Yucatan litters (70%) independent of the transgene, whereas gallbladder and gonad malformations were more frequent in various litters with the same transgene. These results show that the use of cloning in pigs results in a considerable loss of piglets due to malformations and transgenic transformation of the cells used for cloning superimpose on this problem. In combination, these elements could seriously compromise the use of pigs as a model for human diseases and the choice of breeds and also transgenes for this kind of work should be considered carefully. However, further improvements in production of cloned/transgenic embryos may ultimately reduce the incidence of malformations. Table 1.Number of malformations in 54 litters of cloned or transgenic piglets