Effect of donor cell age on the efficiency of nuclear transfer in rabbits

Abstract

The ability of rabbit fibroblasts of different ages to be reprogrammed following nuclear transfer (NT) to aged recipient oocytes was evaluated. The rate of NT blastocysts reconstructed with presumptive G1 stage morula cells or fetal fibroblasts was significantly higher (41.5% and 51.4%) than was those of cloned embryos reconstructed with fibroblasts from young (4-month-old) or aged (5-year-old) animals (16.7% and 7.1%, respectively, P < 0.025). Serum starvation significantly increased the development of NT embryos to the morula-blastocyst stage (67.6% versus 22.9%, P < 0.025). Transfer of 168 NT embryos derived from nuclei of morula cells and 106 control embryos into 21 recipients resulted in 10 pregnancies, 2 NT and 18 control pups, respectively. In the first experiment, transfer of 142 cleaved NT embryos reconstructed with fetal fibroblasts and 86 control embryos into eight recipient does resulted in five pregnancies and the birth of 20 control pups. In the second experiment, after transfer of 112 NT embryos derived from fetal fibroblasts into six recipients, 10 (8.9%) sites of implantation were revealed in two does (33.3%) on day 14 of gestation. This study provides evidence that nuclei of morula cells and fetal and adult fibroblasts differ in their ability to be reprogrammed by recipient cytoplasm following nuclear transfer.