Liu Haijun, Peng Hui, Liu Fang, Ma Qun, Zhang Wenchang
Tianjin Institute of Animal Science and Veterinary Medicine, Tianjin, 300384, China.
College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, China.
In Vitro Cell Dev Biol Anim. 2016 May;52(5):555-61. doi: 10.1007/s11626-015-9984-x. Epub 2016 Mar 4.
The present study aimed to detect the expression of β-galactosidase during long-term cultured goat skin fibroblasts and investigate the effects of donor goat age, sex, and cell passage on senescence and the effects of donor cell passage on in vitro development of nuclear transfer embryos. The results showed that, in the same cell passage, more β-galactosidase-positive cells were detected in cells from older donors than younger donors. Irrespective of the donor age, the number of positive cells was higher in later passages from passages 20 to 50. In the same passage from 20 to 50, the β-galactosidase-positive rate was higher in cells from 5-yr female goat than 5-yr male goat. Using fibroblasts from male goats at various passages as donor cells, reconstructed embryos had similar fusion and cleavage rates, but the blastocyst rate was higher for cells at passages 10 and 20 than passage 30. In conclusion, donor goat age and cell passage had significant effects on the β-galactosidase-positive rate; also, cells from 5-yr female goat had a higher β-galactosidase-positive rate than those from 5-yr male goat, and the donor cell passage affected the developmental potential of nuclear transfer embryos.
本研究旨在检测长期培养的山羊皮肤成纤维细胞中β-半乳糖苷酶的表达情况，并研究供体山羊年龄、性别和细胞传代对细胞衰老的影响，以及供体细胞传代对核移植胚胎体外发育的影响。结果表明，在相同细胞传代次数下，来自老年供体的细胞中检测到的β-半乳糖苷酶阳性细胞比年轻供体的更多。无论供体年龄如何，在第20至50代的后期传代中阳性细胞数量更高。在第20至50代的相同传代中，5岁雌性山羊的细胞中β-半乳糖苷酶阳性率高于5岁雄性山羊。以不同传代的雄性山羊成纤维细胞作为供体细胞，重构胚胎的融合率和分裂率相似，但第10代和第20代细胞的囊胚率高于第30代。总之，供体山羊年龄和细胞传代对β-半乳糖苷酶阳性率有显著影响；此外，5岁雌性山羊的细胞β-半乳糖苷酶阳性率高于5岁雄性山羊，且供体细胞传代影响核移植胚胎的发育潜能。
