Clinical experience with embryos produced by handmade cloning: work in progress.

ABSTRACT

Nuclear transfer in cattle has been shown to cause a high frequency of conceptus loss, excessive accumulation of allantoic fluid, increased birth weight as well as peri- and neonatal deaths. The aims of this preliminary study were to investigate the in vivo development of embryos and fetuses produced by a novel somatic cell cloning method, denominated handmade cloning (HMC), and to characterize the premature calves delivered by Caesarian section. Twenty-five day 7 fresh embryos including seven blastocysts produced by aggregation of two day 4 embryos, and seven vitrified embryos were transferred to synchronized Holstein-Friesian heifers. Embryos produced by aggregation had higher in vivo developmental competence than single embryos (67% versus 38% pregnancy rate on day 28). On days 28, 42, 63 and 250 after estrus, 12 (48%), 5 (20%), 3 (12%) and 2 (8%) recipients of fresh embryos remained pregnant, while 1 recipient of a vitrified embryo was pregnant. One recipient was euthanized due to development of hydrallantois. Caesarian sections were performed on the remaining three recipients on days 252 or 259. The premature calves weighed 60 kg, 47 kg and 45 kg, respectively, and displayed increased weights of body, heart, liver, kidneys, thyroid glands and increased size of placentomes. Furthermore, they had reduced respiratory function, hypoxia, acidosis and altered glucose metabolism. In conclusion, these preliminary data show that handmade somatic cell cloning resulted in an overall delivery rate of 9%, one case of hydrallantois (3%), oversized placentomes and fetuses, disproportionate growth of several internal organs and metabolic immaturity of the premature calves.