Effects of SOF and CR1 media on developmental competence and cell apoptosis of ovine in vitro fertilization embryos.
The present study was to investigate effects of synthetic oviductal fluid (SOF) and Charles Rosenkrans medium (CR1) culture systems on developmental competence and cell apoptosis of ovine in vitro fertilization (IVF) embryos. Ovine presumptive IVF zygotes were cultured in the following six media: (1) SOF supplemented with amino acids (SOFaa) and 8 mg/ml bovine serum albumin (BSA) for 9 days (SOFaaBSA); (2) SOFaa supplemented with 10% fetal bovine serum (FBS) for 9 days (SOFaaFBS); (3) SOFaaBSA for first 3 days and then SOFaaFBS for later 6 days (SOFaaBSA-FBS); (4) CR1 supplemented with amino acids (CR1aa) and 8 mg/ml BSA for 9 days (CR1aaBSA); (5) CR1aa supplemented with 10% FBS for 9 days (CR1aaFBS); (6) CR1aaBSA for first 3 days and then CR1aaFBS for later 6 days (CR1aaBSA-FBS). The rates of blastocyst and hatched blastocyst in group 1, group 3 and group 6 were not different (P>0.05), but were greater than in other three groups (P<0.05). In SOF and CR1 cultural system, SOFaaBSA and CR1aaBSA-FBS provided the highest blastocyst rates respectively. Both numbers of total cell and trophectoderm (TE) in expanded or hatched blastocyst from SOFaaBSA were significantly higher than CR1aaBSA-FBS (P<0.05). However, the inner cell mass (ICM) cell number and ratio of ICM to TE cell in expanded or hatched blastocysts were not different between two groups (P>0.05). The apoptotic signals were firstly observed at 8-cell stage in two groups and became stronger and stronger with the development of embryos. Rates of embryos with apoptotic signals in group 6 at morula or blastocyst were greater than in group 1 (P<0.05). The apoptotic nuclei numbers of morula or blastocyst in group 6 were also significantly higher than group 1 (P<0.05). It is concluded that CR1aaBSA-FBS can support in vitro development of ovine IVF embryos, but SOFaaBSA is more suitable.
Wan
Peng-cheng
PC
Laboratory of Animal Embryonic Biotechnology, College of Animal Science and Technology, China Agricultural University, Beijing, PR China.
Hao
Ze-dong
ZD
Zhou
Ping
P
Wu
Yi
Y
Yang
Ling
L
Cui
Mao-sheng
MS
Liu
Shou-ren
SR
Zeng
Shen-ming
SM
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002452:Cell Count; D003470:Culture Media / Q000737:chemistry*; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary*; D012756:Sheep / Q000196:embryology*; D013194:Staining and Labeling; D015053:Zygote / Q000254:growth & development*
D003470:Culture Media