Peura T T, Hartwich K M, Hamilton H M, Walker S K
South Australian Research and Development Institute, Turretfield Research Centre, Rosedale, SA, Australia.
Reprod Fertil Dev. 2003;15(3):157-65. doi: 10.1071/rd02092.
The aim of this study was to compare serum-starved and non-starved donor cells in sheep nuclear transfer with a special emphasis on cloning outcomes. Sheep oocytes, derived either in vivo or in vitro, were fused with cultured serum-starved or actively growing adult granulosa cells. Resulting blastocysts were transferred to recipients fresh or after vitrification, and subsequent pregnancies followed to term. Donor cell treatment did not significantly affect preimplantation development, pregnancy rates, fetal loss or neonate survival rates. Of 22 lambs born, ten survived the immediate perinatal period but all succumbed at various timepoints within the first few weeks of life. The results of the study suggest that the use of serum-starved cells offers no advantages or disadvantages to cloning outcomes. Neither were significant differences in outcomes observed when using either in vivo- or in vitro-derived oocytes or embryos transferred fresh or after vitrification. Yet, these results continue to highlight problems associated with somatic cell cloning as indicated by offspring mortality. It remains unclear whether the high offspring mortality in the current study was related to species, associated with the cell lines used or the result of other causes.
本研究的目的是比较绵羊核移植中血清饥饿和未饥饿的供体细胞,特别关注克隆结果。体内或体外获得的绵羊卵母细胞与培养的血清饥饿或活跃生长的成年颗粒细胞融合。将所得囊胚新鲜移植或玻璃化后移植给受体,并追踪随后的妊娠至足月。供体细胞处理对植入前发育、妊娠率、胎儿丢失或新生儿存活率没有显著影响。在出生的22只羔羊中,10只在围产期即刻存活,但全部在出生后几周内的不同时间点死亡。该研究结果表明,使用血清饥饿细胞对克隆结果没有优势或劣势。使用新鲜或玻璃化后的体内或体外获得的卵母细胞或胚胎时,在结果上也未观察到显著差异。然而,这些结果继续突出了体细胞克隆相关的问题,如后代死亡率所示。目前研究中高后代死亡率是与物种有关、与所用细胞系有关还是其他原因的结果仍不清楚。