Abstract
Experiments were conducted to evaluate the effect of blastomere diameters and cell cycle stages on the subsequent development of nuclear transplant rabbit embryos (NT-embryos) using nuclei derived from the 16- or 32-cell stage embryos. All blastomeres and NT-embryos were cultured individually in modified Ham's F-10 medium supplemented with 10% rabbit serum (RS) at 38°C and 5% CO 2 in air. The diameter of blastomeres from 16-cell stage embryos was found twice of those from 32-cell stage (51 vs 27 μm). Significant differences were observed in cleavage rates (>3 divisions) in the isolated single blastomeres (54 vs 48 for 16-cell; 28 vs 14 for 32-cell, p 3 divisions) of the NT-embryos were greater in the small nuclear donors than in the large donors (73 vs 55%, p 0.05). In conclusion, it appeared that no difference in the developmental competence between large and small isolated blastomeres was observed. When smaller 16-cell stage blastomeres were used as nuclear donor, the cleavage rate or development of NT-embryos was improved and was compromised when 32-cell stage blastomeres were used. Therefore, control nuclear stage of the donor cell at G 1 phase in preactivated nuclear recipients seemed to be beneficial for the cleavage rate of the reconstructed embryo in the 16-cell transplant, but not for subsequent morula or blastocyst development.
Highlights
Nuclear cloning is a powerful tool for large-scale production of identical multiplets containing the same genetic materials
Large and small blastomeres from either 16- or 32-cell embryos were transferred into the recipient oocytes
The reconstructed embryos were cultured in Ham's F-10
Summary
Nuclear cloning is a powerful tool for large-scale production of identical multiplets containing the same genetic materials. A German developmental biologist, in 1938, this technology was demonstrated by Briggs and King (1952) in Xenopus oocytes for the first time. Solter et al (1980) applied this technology to mammalian species and compiled data including nuclear transplant mice, rabbits, pigs, sheep, goats, cows (Wolf et al, 1999) and Rhesus monkey (Meng et al, 1997) were generated thereafter. A milestone breakthrough with the birth of Dolly the sheep was achieved by Wilmut et al (1997) using adult mammary gland cells as the donor nucleus. Cloned goats (Baguisi et al, 1999), cattle (Cibelli et al, 1998; Wells et al, 1999; Zakhartchenko et al, 1999; Kubota et al, 2000), and pigs.
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