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BOR - Papers in Press, published online ahead of print December 15, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.035105
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BIOLOGY OF REPRODUCTION 72, 932–936 (2005)
DOI: 10.1095/biolreprod.104.035105
© 2005 by the Society for the Study of Reproduction, Inc.

Establishment of Male and Female Nuclear Transfer Embryonic Stem Cell Lines from Different Mouse Strains and Tissues1

Sayaka Wakayama3,4,, Hiroshi Ohta3, Satoshi Kishigami3, Nguyen Van Thuan3, Takafusa Hikichi3, Eiji Mizutani3,5,, Masashi Miyake4, and Teruhiko Wakayama2,3

Center for Developmental Biology,3 RIKEN Kobe, Kobe 650-0047, Japan Department of Life Science,4 Graduate School of Science and Technology, Kobe University, Kobe 657-8501, Japan Graduate School of Agricultural Science,5 Tohoku University, Sendai 981-8555, Japan

Nuclear transfer can be used to generate embryonic stem cell lines from somatic cells, and these have great potential in regenerative medicine. However, it is still unclear whether any individual or cell type can be used to generate such lines. Here, we tested seven different male and female mouse genotypes and three cell types as sources of nuclei to determine the efficiency of establishing nuclear transfer embryonic stem cell lines. Lines were successfully established from all sources. Cumulus cell nuclei from F1 mouse genotypes showed a significantly higher cumulative establishment rate from reconstructed oocytes than from other cells; however, there were no genotype differences in success rates from cloned blastocysts. Thus, the overall success depends on preimplantation development, and, once embryos have reached the blastocyst stage, the genotype differences disappear. All mouse genotypes that were tested demonstrated at least one cell line that subsequently contributed to germline transmission in chimeric mice, so these cell lines clearly possess the same potential as embryonic stem cells derived from fertilized embryos. Thus, nuclear transfer embryonic stem cells can be generated relatively easily from a variety of inbred mouse genotypes and cell types of both sexes, even though it may be more difficult to generate clones directly.

1 Supported by a Grant-in-Aid for Creative Scientific Research (13GS0008), Scientific Research in Priority Areas (15080211), Young Scientists A (15681014), and a project for the realization of regenerative medicine (the research field for the technical development of stem cell manipulation) to T.W. from the Ministry of Education, Science, Sports, Culture and Technology of Japan.

2 Correspondence: Teruhiko Wakayama, 2-2-3 Minatojima-minamimachi Chuo-ku, Kobe 650-0047, Japan. FAX: 81 78 306 0101;teru{at}cdb.riken.go.jp






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Copyright © 2005 by the Society for the Study of Reproduction.