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Regarding cloned animals, interesting questions have been raised as to whether cloning restores cellular senescence undergone by their donor cells and how long cloned animals will be able to live. Focusing our attention on differences in telomere lengths depending on the tissue, we had produced 14 cloned cattle by using nuclei of donor cells derived from muscle, oviduct, mammary, and ear skin. Here, we show remarkable variation in telomere lengths among them using Southern blot analysis with telomere-specific probe. Telomere lengths in cloned cattle derived from muscle cells of an old bull were longer than those of a donor animal but were within the variation in normal calves. On the other hand, those derived from oviductal and mammary epithelial cells of an equally old cow were surprisingly shorter than any found in control cattle. The telomere lengths of cloned cattle derived from fibroblasts and oviductal epithelial cells of younger cattle showed the former and the latter results, respectively. In both cases, however, less telomere erosion or telomere extension from nuclear transfer to birth in most cloned cattle was observed in comparison with telomere erosion from fertilization to birth in control cattle. Embryonic cell-cloned cattle and their offspring calves were also shown to have telomeres longer than those in age-matched controls. These observations indicate that cloning does not necessarily restore the telomere clock but, rather, that nuclear transfer itself may commonly trigger an elongation of telomeres, probably more or less according to donor cell type. Remarkable variations among cloned cattle are suggested to be caused by variation in telomere length among donor cells and more or less elongation of telomere lengths induced by cloning.

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We previously showed that telomere lengths of 10 somatic cell cloned cows were significantly shorter than normal. In this study, we investigated growth, reproduction, and lactation in these animals to determine if shortened telomeres have any effect on these characteristics. Six Holstein and 4 Jersey cloned cows, derived from oviduct cells, were reared under general group feeding. Body weights were recorded from birth to 48 mo of age. A number of reproductive characteristics were screened during the prepubertal, postpubertal, and postpartum periods. After parturition, milk yields were recorded daily and percentages of milk fat, proteins, and solids-not-fat were measured at monthly intervals. These data were used to estimate production of milk components over a 305-d period. Overall, the cloned heifers exceeded standard growth rates for each breed. The cows were inseminated at the first estrus after they reached 450 d of age, and delivered normal calves except for one stillbirth in the Holstein group. They were inseminated at postpartum estrus to provide second and third parturitions and, again, these pregnancies were normal. Gestational periods and birth weights of the calves were both within the normal range. The average total milk yield per cow in Holstein group clones was less than that of the original cow, whereas Jersey group clones showed a higher average milk yield than the original cow. In both groups of cloned cows, inter-individual variation in milk production was relatively large; however, the coefficient of variation was less than 10%. Our results suggest that the cloned cows have normal growth, reproductive, and lactation characteristics, and thus normal productivity, despite having reduced telomere lengths.

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