The differential requirement of albumin and sodium citrate on the development of in vitro produced bovine embryos.
In vitro culture for bovine embryos is largely not optimal. Our study was to determine the components necessary for early embryo development. In experiment 1, IVF embryos were cultured for two days in CR1aa medium containing sodium citrate and BSA from two sources (Sigma vs. ICPbio), subsequently for additional five days with cumulus monolayer in 10% FBS CR1aa. We found that supplementation with both Sigma-BSA and sodium citrate significantly increased total blastocyst (BL) development compared with the ICPbio-BSA groups (37% vs. 19-21%), and enhanced the total number of high quality (C1 BL, IETS standard) blastocysts (26% vs. 11-17%) (P < 0.05). In experiment 2 with serum free and/or somatic free culture, we found that CR1aa culture can support a comparable embryo development with a supplement of Sigma BSA. The addition of sodium citrate did not increase blastocyst development in either the Sigma-BSA or the ICPbio-BSA groups. An inferior blastocyst development occurring in ICPbio-BSA culture (1-3%) could be rescued by culture in CRlaa supplemented with 10% FBS (29%), more importantly, by culture in CR1aa with a replacement of Sigma BSA (24%) (P <0.05). C1 blastocysts rescued by FBS and Sigma BSA in ICPbio-BSA culture possessed indistinguishable morphology to embryos developed in a Sigma-BSA, FBS and somatic co-culture system, showing similar cell number/blastocyst (129-180, P > 0.05). Our study found a beneficial effect of sodium citrate and BSA on the in vitro development of bovine IVF embryos during co-culture. We also determined that differential embryotrophic factor(s) contained in BSA and serum, probably not sodium citrate, is necessary for promoting competent morula and blastocyst development in cattle.
Sung
Li-Ying
LY
Department of Animal Science and Connecticut Center for Regenerative Biology, University of Connecticut, Storrs, CT 06269, USA.
Du
Fuliang
F
Xu
Jie
J
Chang
Wancun
W
Nedambale
Tshimangadzo Lucky
TL
Zhang
Jiaxin
J
Jiang
Shie
S
Tian
X Cindy
XC
Yang
Xiangzhong
X
D000418:Albumins / Q000008:administration & dosage*; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000196:embryology*; D002951:Citrates / Q000008:administration & dosage*; D018920:Coculture Techniques / Q000662:veterinary; D003470:Culture Media; D046149:Embryo Culture Techniques / Q000662:veterinary*; D047108:Embryonic Development / Q000502:physiology*; D005307:Fertilization in Vitro / Q000662:veterinary; D009751:Nutritional Requirements; D012710:Serum Albumin, Bovine / Q000008:administration & dosage; D000077559:Sodium Citrate
D000418:Albumins; D002951:Citrates; D003470:Culture Media; D000077559:Sodium Citrate; D012710:Serum Albumin, Bovine