In vitro production of bovine embryos in medium supplemented with a serum replacer: effects on blastocyst development, cryotolerance and survival to term.
In this study, we evaluated a serum replacer (SR; Knockout SR, Invitrogen) in our in vitro culture systems. We hypothesized that SR would benefit bovine embryo development, since SR supported survival of embryonic stem cells (which originate from embryos). Experiment 1 compared oocyte maturation with SR versus fetal bovine serum (FBS). Following fertilization, blastocyst development was lower for oocytes matured with SR (21.5 versus 34.1, P<0.05). Experiment 2 evaluated SR for culturing embryos. Following fertilization, embryos were cultured for 3 days in KSOM, and then assigned to treatments: (1) KSOM static culture (KNM); (2) fresh KSOM (KD3); (3) KSOM+SR or (4) KSOM+FBS and cultured to Day 7 (fertilization=Day 0). Blastocyst development in FBS or SR was higher than either KNM or KD3 (48.2, 47.2, 32.7, and 35.5, respectively, P<0.05). Experiment 3 evaluated cryosurvival of embryos cultured in the same manner as Experiment 2. On Day 7, embryos were vitrified and upon warming, embryos cultured in SR had greater 24h survival rates (70.6%) than all other treatments (P<0.05). Finally, Experiment 4 evaluated effects of SR on pregnancy rate and development to term. Culture in SR was not detrimental to pregnancy or calving rates (50 and 50%, respectively), and SR calves had normal birth weights (mean=38.8 kg+/-1.5). In conclusion, the use of SR for maturation of oocytes was not beneficial; however, SR enhanced embryo culture by improving development in vitro, cryotolerance and survival, effectively replacing serum in culture.
Moore
K
K
Department of Animal Sciences, University of Florida, Gainesville, FL 32611-0910, USA. kmoore@animal.ufl.edu
Rodríguez-Sallaberry
C J
CJ
Kramer
J M
JM
Johnson
S
S
Wroclawska
E
E
Goicoa
S
S
Niasari-Naslaji
A
A
D000818:Animals; D001724:Birth Weight / Q000187:drug effects; D002417:Cattle / Q000196:embryology* / Q000502:physiology; D015925:Cryopreservation / Q000662:veterinary*; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000502:physiology*; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005306:Fertilization / Q000187:drug effects; D005307:Fertilization in Vitro / Q000662:veterinary; D008297:Male; D011247:Pregnancy; D018873:Pregnancy Rate; D044967:Serum / Q000502:physiology; D012744:Sex Ratio; D016019:Survival Analysis
D003470:Culture Media