Effect of the timing of the first cleavage on the developmental potential of nuclear-transferred mouse oocytes receiving embryonic stem cells.
The present study examined whether the timing of the first cleavage has an effect on the in vitro and in vivo developmental potential of nuclear-transferred mouse oocytes receiving embryonic stem cells. First, the timing of the first cleavage and the developmental potential of nuclear-transferred oocytes were examined every hour from 12 to 24 h after the start of culture and compared with in vitro-fertilized oocytes. The developmental potential of in vitro-fertilized oocytes decreased gradually according to the time required for cleavage (84% (32/38) for 15 h to 50% (1/2) for 20 h), but intermediate-cleaved (15-16 h) nuclear-transferred oocytes had a higher potential to develop into blastocysts (55% (17/31) to 67% (45/67) versus 0-43% (6/14)]. Second the nuclear-transferred oocytes were divided into three groups according to the timing of the first cleavage; each group was cultured to blastocysts in vitro, and then transferred to recipients. The potential of intermediate-cleaved oocytes (15-16 h) to develop into blastocysts was significantly higher than fast-cleaved (before 15 h) and slow-cleaved (after 16 h) oocytes (65, 46, and 37%). The proportion of fetuses on Day 10.5 of pregnancy was highest in the intermediate-cleaved group (4 versus 2 and 1%, respectively) and a full-term fetus was obtained from this group. The present study demonstrated that the timing of the first cleavage could be used to determine the potential of nuclear-transferred oocytes with embryonic stem cells to develop to the blastocyst stage in vitro, but not to determine post-implantation viability after transfer to recipients.
Kobayashi
T
T
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, 3327-204, Nakamachi, Nara 631-8505, Japan.
Kato
Y
Y
Tsunoda
Y
Y
D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002970:Cleavage Stage, Ovum; D019976:Cloning, Organism; D046508:Culture Techniques; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D005307:Fertilization in Vitro; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000502:physiology / Q000648:ultrastructure*; D011247:Pregnancy; D013234:Stem Cells / Q000648:ultrastructure*; D013997:Time Factors