Production of cloned mice from embryonic stem cells arrested at metaphase

In mammals, cloned individuals can be produced from somatic cells. The combined use of gene targeting in embryonic stem cells and cloning contributes to the investigation of gene function in mammals. However, one of the major limitations to cloning is the low viability of cloned embryos, leading typically to high rates of pre- and postnatal death. The present study investigated whether cloning efficiency is influenced by the procedural differences involved in using transfected embryonic stem cells arrested at M phase for cloning by both single and serial transfer. In contrast to a previous study, in which fibroblasts were used, in the present study using embryonic stem cells there was no difference in the rate of production of cloned pups after the use of a single or serial nuclear transfer, although the proportion of blastocysts (70% versus 51%) was significantly higher (P < 0.001) after serial nuclear transfer. After embryo transfer of 445 blastocysts, 218 (49%) implanted and 27 (6% of blastocysts transferred) live pups were born. Of these 27 pups, 23 developed to adults of apparently normal fertility. Of these adults, 39% (n = 9) were derived from targeted embryonic stem cells, which is similar to the proportion of targeted embryonic stem cells in the population used for cloning. This study showed that cloning with embryonic stem cells is a viable procedure resulting in the production of transgenic cloned adults.

This article has been cited by other articles:

				C. Palmieri, P. Loi, G. Ptak, and L.D. Salda REVIEW PAPER: A Review of the Pathology of Abnormal Placentae of Somatic Cell Nuclear Transfer Clone Pregnancies in Cattle, Sheep, and Mice Vet. Pathol., November 1, 2008; 45(6): 865 - 880. [Abstract] [Full Text] [PDF]

				Y. Jincho, Y. Sotomaru, M. Kawahara, Y. Ono, H. Ogawa, Y. Obata, and T. Kono Identification of Genes Aberrantly Expressed in Mouse Embryonic Stem Cell-Cloned Blastocysts Biol Reprod, April 1, 2008; 78(4): 568 - 576. [Abstract] [Full Text] [PDF]

				K. Inoue, S. Noda, N. Ogonuki, H. Miki, S. Inoue, K. Katayama, K. Mekada, H. Miyoshi, and A. Ogura Differential Developmental Ability of Embryos Cloned from Tissue-Specific Stem Cells Stem Cells, May 1, 2007; 25(5): 1279 - 1285. [Abstract] [Full Text] [PDF]

				A. Schurmann, D. N Wells, and B. Oback Early zygotes are suitable recipients for bovine somatic nuclear transfer and result in cloned offspring. Reproduction, December 1, 2006; 132(6): 839 - 848. [Abstract] [Full Text] [PDF]

				N. Shimozawa, Y. Sotomaru, N. Eguchi, S. Suzuki, K. Hioki, T. Usui, T. Kono, and M. Ito Phenotypic abnormalities observed in aged cloned mice from embryonic stem cells after long-term maintenance. Reproduction, September 1, 2006; 132(3): 435 - 441. [Abstract] [Full Text] [PDF]

				D. R Arnold, V. Bordignon, R. Lefebvre, B. D Murphy, and L. C Smith Somatic cell nuclear transfer alters peri-implantation trophoblast differentiation in bovine embryos. Reproduction, August 1, 2006; 132(2): 279 - 290. [Abstract] [Full Text] [PDF]

				Y. Ono and T. Kono Irreversible Barrier to the Reprogramming of Donor Cells in Cloning with Mouse Embryos and Embryonic Stem Cells Biol Reprod, August 1, 2006; 75(2): 210 - 216. [Abstract] [Full Text] [PDF]

				S. Gao, M. McGarry, T. Ferrier, B. Pallante, B. Gasparrini, J. Fletcher, L. Harkness, P. De Sousa, J. McWhir, and I. Wilmut Effect of Cell Confluence on Production of Cloned Mice Using an Inbred Embryonic Stem Cell Line Biol Reprod, February 1, 2003; 68(2): 595 - 603. [Abstract] [Full Text] [PDF]