Laboratory for Genomic Reprogramming
Research Areas
| Cloning mammals by nuclear transfer began over fifteen years ago, yet the basic biology underlying this process remains unclear. We have succeeded in generating cloned mice from adult/fetus somatic and cultured (e.g., ES) cells. As in other mammals, the overall efficiency of mouse cloning in all reports is less than 5%. Why is this efficiency so low? Perhaps only a small percentage of cells in any given population are "cloning-competent" or because of technical problems (e.g., nuclear damage), but there exists a bewildering range of other possibilities. The concept of "nuclear reprogramming" links the efficiency of cloning to changes that an incoming nucleus must undergo to direct development, but its mechanisms are unknown. We have derived ES cell lines via nuclear transfer (ntES cell lines) from adult mouse somatic cells (e.g., tail-tip fibroblasts). ntES cells contribute to an extensive variety of cell types, including germ cells in vivo, suggesting that they have been effectively reprogrammed and are fully pluripotent. Thus, we now have a uniquely powerful research model to investigate the biology of mammalian cloning, including the mechanisms that underlie reprogramming. This is expected to provide insights that will allow us to increase cloning efficiency significantly. |  Team Leader Teruhiko WAKAYAMA (Ph.D.)  |
Research Subjects
| (1) | Improvement of nuclear transfer techniques |
| (2) | Study of genomic reprogramming in mouse oocytes cytoplasm |
| (3) | Comparison with ntES cell line and ES cell line |
List of Selected Publications
| (1) | Ono, T., Mizutani, E., Li, C., and Wakayama, T.: "Nuclear transfer preserves the nuclear genome of freeze-dried mouse cells." J. Reprod Dev. 54: 486-491 (2008) |
| (2) | Bui, H.T., Wakayama, S., Kishigami, S., Thuan, N.V., and Wakayama, T.: "The cytoplasm of mouse germinal vesicle (GV) stage oocytes can enhance somatic cell nuclear reprogramming." Development 135: 3935-3945 (2008) |
| (3) | Wakayama, S., Ohta, H., Hikichi, T., Mizutani, E., Iwaki, T., Kanagawa, O., and Wakayama, T.: "Production of healthy cloned mice from bodies frozen at -20oC for 16 years." Proc Natl Acad Sci USA. 105, 17318-17322(2008) |
| (4) | Tabar, V., Tomishima, M., Panagiotakos, G., Wakayama, S., Menon, J., Chan, B., Mizutani, E., Al-Shamy, G., Ohta, H., Wakayama, T., and Studer, L.: "Therapeutic cloning in individual parkinsonian mice." Nat. Med. 14: 379-381 (2008) |
| (5) | Ohta, H., Sakaide, Y., and Wakayama, T.: "The Birth of Mice from Testicular Spermatozoa Retrieved from Frozen Testicular Sections." Biol. Reprod. (2008) in press |
| (6) | Ohta, H., Sakaide, Y., and Wakayama, T.: "Generation of progeny via ICSI following enrichment of elongated spermatids from mouse testis by flow-cytometric cell sorting." Hum. Reprod. 22: 1612-1616(2007) |
| (7) | Wakayama, S., Suetsugu, R., Thuan, NV., Ohta, H., Kishigami, S., and Wakayama, T.: "Establishment of mouse embryonic stem cell lines from somatic cell nuclei by nuclear transfer into aged, fertilization-failure mouse oocytes." Curr. Biol. 17: R120-R121 (2007). |
| (8) | Wakayama, S., Hikichi, T., Suetsugu, R., Sakaide, Y., Bui, H.T., Mizutani, E., and Wakayama, T.: "Efficient establishment of mouse embryonic stem cell lines from single blastomeres and polar bodies." Stem Cells. 25:986-993 (2007) |
| (9) | Hikichi, T., Wakayama, S., Mizutani, E., Takashima, Y., Kishigami, S., Van Thuan, N., Ohta, H., Bui, HT., Nishikawa, SI., and Wakayama, T.: "Differentiation Potential of Parthenogenetic Embryonic Stem Cells is Improved by Nuclear Transfer." Stem Cells. 25:46-53 (2007) |
| (10) | Mizutani, E., Ohta, H., Kishigami, S., Thuan, N.V., Hikichi, T, Wakayama, S., Kosaka, M., Sato, E., and Wakayama, T.: "Developmental ability of cloned embryos from neural stem cells." Reproduction 132:849-857 (2006) |