title: In vitro and in vivo developmental potential of nuclear transfer embryos using bovine cumulus cells prepared in four different conditions.
abstract: We examined the effect of culture of donor cells on nuclear transfer efficiency using bovine cumulus cells treated with four different conditions: (1). group A, the cells removed from cumulus-oocyte complexes (COC) after aspiration of ovarian follicles; (2). group B, the cells removed from COC after in vitro maturation; (3). group C, the cells cultured in Dulbecco's Modified Eagle's Medium (DMEM) with 10% fetal bovine serum (FBS) for 3 days after some subculture; and (4). group D, the cells cultured in DMEM with 0.5% FBS for an additional 5 days. Analysis of cell cycle using flow cytometry revealed that the relative proportion of donor cells at G0/G1 phase of cell cycle was 89.7% in group A, 89.5% in group B, 76.0% in group C, and 90.6% in group D. The developmental rates to blastocyst stage in groups C (45.3%) and D (46.4%) were significantly (p < 0.05) higher than in groups A (17.5%) and B (31.9%). After transfer of blastocysts produced in each group, nine of 24 recipients became pregnant on day 30. A total of five live calves were obtained from cumulus cells in all groups (group A [n = 1], group B [n = 1], group C [n = 2], and group D [n = 1]).
authors.affiliation: Department of Animal Breeding and Reproduction, National Institute of Livestock and Grassland Science, Ibaraki, Japan. akagi@affrc.go.jp
authors.forename: Satoshi
authors.initials: S
authors.lastname: Akagi
authors.forename: Seiya
authors.initials: S
authors.lastname: Takahashi
authors.forename: Noritaka
authors.initials: N
authors.lastname: Adachi
authors.forename: Kiyotoshi
authors.initials: K
authors.lastname: Hasegawa
authors.forename: Toru
authors.initials: T
authors.lastname: Sugawara
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Tozuka
authors.forename: Eiji
authors.initials: E
authors.lastname: Yamamoto
authors.forename: Manabu
authors.initials: M
authors.lastname: Shimizu
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Izaike
mesh_terms: D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D017077:Culture Media, Conditioned; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D053652:Nuclear Transfer Techniques; D009865:Oocytes; D006080:Ovarian Follicle / Q000166:cytology; D011247:Pregnancy; D013997:Time Factors
chemical_list: D017077:Culture Media, Conditioned