Production of blastocysts after intergeneric nuclear transfer of goral (Naemorhedus goral) somatic cells into bovine oocytes.
Interspecies cloning may be a useful method to help conserve endangered species and to study nuclear-cytoplasm interaction. The present study investigated in vitro development of goral (Naemorhedus goral) intergeneric nuclear transfer embryos produced by fusing goral fibroblasts with enucleated metaphase II (MII) bovine oocytes. After two to five passages, serum-starved or non-starved goral skin fibroblast cells were transferred into enucleated MII bovine oocytes. Couplets were electrically fused and chemically activated, and then cultured in either modified synthetic oviduct fluid (mSOF) or tissue culture medium-199 (TCM-199) supplemented with 10% FBS. Serum starvation of donor cells did not affect the fusion rate and or development to of cells to the two-cell stage, to more than 9-cells, or to morulae, regardless of culture medium. Three blastocysts from 202 fused embryos were obtained when embryos reconstructed with non- serum- starved donor cells were cultured in mSOF. However, no blastocysts were obtained when the embryos reconstructed with serum-starved donor cells were cultured in mSOF. The total cell number of goral intergeneric embryos averaged 130.3 (range 105-180). In conclusion, this study demonstrated that bovine oocytes can support blastocyst development after intergeneric SCNT with goral fibroblasts.
Oh
Byoung-Chol
BC
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, and The Xenotransplantation Research Center, Seoul National University Hospital, Korea.
Kim
Jung-Tae
JT
Shin
Nam-Shik
NS
Kwon
Soo-Whan
SW
Kang
Sung-Keun
SK
Lee
Byeong-Chun
BC
Hwang
Woo-Suk
WS
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D019976:Cloning, Organism / Q000379:methods / Q000662:veterinary*; D005347:Fibroblasts / Q000502:physiology; D016014:Linear Models; D053652:Nuclear Transfer Techniques / Q000662:veterinary*; D009865:Oocytes / Q000502:physiology; D012418:Ruminants / Q000196:embryology*; D013045:Species Specificity