Abstract
The nucleus of a somatic cell could be dedifferentiated and reprogrammed in an enucleated heterogeneous oocyte. Some reconstructed oocytes could develop into blastocysts in vitro, and a few could develop into term normally after transferred into foster mothers, but most of cloning embryos fail to develop to term. In order to evaluate the efficacy of embryonic stem cell as nucleus donor in interspecific animal cloning, we reconstructed enucleated rabbit oocytes with nuclei from mouse ES cells, and analyzed the developmental ability of reconstructed embryos in vitro. Two kinds of fibroblast cells were used as donor control, one derived from ear skin of an adult Kunming albino mouse, and the other derived from a mouse fetus. Three types of cells were transferred into perivitelline space under zona pellucida of rabbit oocytes respectively. The reconstructed oocytes were fused and activated by electric pulses, and cultured in vitro. The developmental rate of reconstructed oocytes derived from embryonic stem cells was 16.1%, which was significantly higher than that of both the adult mouse fibroblast cells (0%–3.1%, P< 0.05) and fetus mouse fibroblast cells (2.1%–3.7%, P<0.05). Chromosome analysis confirmed that blastocyst cells were derived from ES donor cell. These observations show that reprogramming is easier in interspecific embryos reconstructed with ES cells than that reconstructed with somatic cells, and that ES cells have the higher ability to direct the reconstructed embryos development normally than fibroblast cells.
Similar content being viewed by others
References
Kato, Y., Tani, T., Sotomaru, Y. et al., Eight calves cloned from somatic cells of a single adult, Science, 1998, 282: 2095.
Wilmut, I., Schnieke, A. E., Mcwhir, J. et al., Viable offspring derived from fetal and adult mammalian cells, Nature, 1997, 385: 810.
Eggan, K., Akutsu, H., Loring, J. et al., Hybrid vigor, fetal overgrowth and viability of mice derived by nuclear cloning and tetraploid embryo complementation, Proc. Nat. Acad. Sci., 2001, 98: 6201.
Polejaeva, I. A., Chen, S. H., Vaught, T. D. et al., Cloned pigs produced by nuclear transfer from adult somatic cells, Nature, 2000, 407: 505.
Loi, P., Ptak, G., Barboni, B. et al., Genetic rescue of an endangered mammal by cross-species nuclear transfer using post-mortal somatic cells, Nat. Biotechnol., 2001, 19: 962.
Chen, D. Y., Sun, Q. Y., Liu, J. L. et al., The giant panda (Ailuropoda melanoleuca) somatic nucleus can defferentiate in rabbit ooplasm and support early development of the reconstructed egg, Science in China, Ser. C., 1999, 42: 346.
Dominko, T., Mitalipova, M., Haleg, B. et al., Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species, Biol. Reprod., 1999, 60: 821.
Dean, W., Bowden, L., Aitchison, A. et al., Altered imprinted gene methylation and expression in completely ES cell-derived mouse fetuses: association with aberrant phenotypes, Development, 1998, 125: 2273.
Evans, M. J., Kaufman, M. H., Establishment in culture of pluripotential cell from mouse embryos, Nature, 1981, 292: 154.
Wakayama, T., Rodriguez, I., Perry, A. C. F. et al., Mice cloned from embryonic stem cell, Proc. Nat. Acad. Sci., 1999, 96: 14984.
Robertson, E. J., Teratocarcinomas and Embryonic Stem Cell, 1st. ed., Oxford: IRL Press, 1987, 147.
Rideout, W. M., Wakayama, T., Wutz, A., Generation of mice from wild-type and targeted ES cells by nuclear cloning, Nat. Genet, 2000, 24: 109.
Wakayama, T., Yanagimachi, R., Cloning of male mice from adult tail-tip cells, Nat. Genet., 1999, 22: 127.
Author information
Authors and Affiliations
Corresponding author
Additional information
The authors contributed equally to this work.
About this article
Cite this article
Wang, H., Zheng, R., Xu, Y. et al. Embryonic stem cell as nuclear donor could promote in vitro development of the heterogeneous reconstructed embryo. Chin.Sci.Bull. 47, 1811–1815 (2002). https://doi.org/10.1007/BF03183848
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF03183848