Cloning of calves from various somatic cell types of male and female adult, newborn and fetal cows.

Y Kato, T Tani, Y Tsunoda
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Abstract

Twenty-four calves were cloned from six somatic cell types of female and male adult, newborn and fetal cows. The clones were derived from female cumulus (n = 3), oviduct (n = 2) and uterine (n = 2) cells, female and male skin cells (n = 10), and male ear (n = 5) and liver (n = 2) cells. On the basis of the number of cloned embryos transferred (n = 172) to surrogate cows, the overall rate of success was 14%, but based on the number of surrogate mothers that became pregnant (n = 50), the success rate was 48%. Cell nuclei from uterus, ear and liver cells, which have not been tested previously, developed into newborn calves after nuclear transfer into enucleated oocytes. To date, seven female and six male calves have survived: six of the females were from adult cells (cumulus (n = 3), oviduct (n = 2) and skin (n = 1) cells) and one was from newborn skin cells, whereas the male calves were derived from adult ear cells (n = 3), newborn liver and skin cells (n = 2), and fetal cells (n = 1). Clones derived from adult cells frequently aborted in the later stages of pregnancy and calves developing to term showed a higher number of abnormalities than did those derived from newborn or fetal cells. The telomeric DNA lengths in the ear cells of three male calves cloned from the ear cells of a bull aged 10 years were similar to those of the original bull. However, the telomeric DNA lengths from the white blood cells of the clones, although similar to those in an age-matched control, were shorter than those of the original bull, which indicates that telomeric shortening varies among tissues.

从各种体细胞类型的雄性和雌性成年牛、新生牛和胎牛中克隆小牛。
从6种体细胞类型的雌、雄、成年、新生和胎牛中克隆了24头小牛。克隆来源于雌性积云细胞(n = 3)、输卵管细胞(n = 2)和子宫细胞(n = 2)、雌性和雄性皮肤细胞(n = 10)、雄性耳细胞(n = 5)和肝脏细胞(n = 2)。如果以移植到代孕牛身上的克隆胚胎数量(172个)为标准,总体成功率为14%,但如果以成功怀孕的代孕母亲数量(50个)为标准,成功率为48%。从子宫、耳部和肝细胞中提取的细胞核,在细胞核移植到去核卵母细胞后发育成新生牛犊,这些细胞核以前没有被测试过。迄今为止,已有七只雌性和六只雄性幼崽幸存下来。6的雌性成年细胞(积云(n = 3),输卵管(n = 2)和皮肤(n = 1)细胞)和一个来自新生儿皮肤细胞,而雄性小牛来自成人耳朵细胞(n = 3),新生儿肝脏和皮肤细胞(n = 2),和胎儿细胞克隆(n = 1)。来自成年细胞经常在怀孕晚期流产,小牛发展术语显示更高数量的异常比那些来自新生儿或胎儿细胞。从一头10岁公牛的耳细胞中克隆出的3头雄性小牛的耳细胞端粒DNA长度与原公牛相似。然而,克隆公牛白细胞的端粒DNA长度虽然与年龄匹配的对照组相似,但却比原始公牛短,这表明端粒缩短在不同组织中有所不同。
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