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ARS Home » Research » Publications at this Location » Publication #85021
Title: SERA SUPPLEMENTS AND BSA EFFECT ON OPTIMIZATION OF RABBIT EMBRYO CULTURE SYSTEMS

Author
item JOHNSON, J - ALABAMA A&M UNIVERSITY
item Johnson, Lawrence
item BRACKETT, B - UNIVERSITY OF GEORGIA
item Dobrinsky, John
item PURSEL, V - USDDA, ARS, LPSI, GEML

Submitted to: Theriogenology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/12/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: Biotechnology research programs require the establishment of procedures that can be used to plan and conduct experiments to gain new knowledge. There is also the necessity for a consistent use of these techniques for training purposes in the classroom and laboratory. These studies were conducted to compare culture media for development of rabbit embryos to blastocysts and to ascertain the most appropriate media for cryobanking. Fetal calf serum or ovine serum gave improved survival rates in comparison to BSA containing media. These results provide a basis for implementing protocols for storing morula and blastocyst stage embryos in liquid nitrogen for long term cryobanking. These results help to establish a standardized protocol for the biotechnology program.

Technical Abstract: The aim of this study was to determine the efficacy of Earles Balanced Salt Solution (EBSS) and Hams F10 (MHF10) to sustain viability of rabbit embryos before and after cryopreservation. Morula stage rabbit embryos (n=339) were cultured in vitro 37C, 5% CO2 in humidified air) in EBBS or MHF10 medium supplemented with 15% fetal calf serum (FCS) or 15% ovine serum (OS) )or 1.5% Bovine Serum Albumin (15 mg/ml, BSA) and compared in 3 x 2 factorial design. Embryo survival was assessed by their ability to develop to blastocysts, and hatched blastocysts, during 24 thru 120 hrs culture. Few embryos (31%) cultured in EBSS alone, developed to blastocysts compared to (86%) achieving blastocyst expansion in MHF10. The addition of 1.5% BSA enhanced the efficacy of EBSS & MHF10 to support blastocyst development 84% and 90%) respectively), but viability was diminished and fewer embryos (17% & 58%, respectively) reached the hatching blastocyst stage. Optimal growth hand expansion of blastocysts occurred in either EBSS or MHF10 supplemented with FCS (91% & 96%) or OS (92% &93%) however, the efficacy of these media shifted in their ability to meet nutrient requirements of hatching embryos (78%, 92%, 45%, 73%, respectively). The data suggests that FCS or OS would be superior to BSA in either EBSS or NHF10 for short term culture of morula to blastocysts prior to embryo cryopreservation or transfer. Long-term cultured benefit using MHF10 + FCS which in this study, maintained support of 41-50% of all embryos cultured to completely hatched blastocysts.