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Effects of Serum on Swine Morulae and Blastocysts in Vitro¹

Kansas State University^,3, Manhattan 66506

Abstract

The effects of serum on development of swine embryos in vitro and the relationship between embryo age and requirements for in vitro growth were investigated. Embryos were cultured either in a modified Krebs-Ringer bicarbonate (mKRB) medium containing glucose and bovine serum albumin or in mKRB supplemented with serum. The addition of 10% heat-treated sheep serum improved development, as indicated by an increase in nuclei (P<.05) and increased survival (P<.10) of day 8 blastocysts cultured in medium containing serum. Blastocysts were classified as surviving if they remained expanded and had a smooth appearance. More (P<.05) blastocysts cultured in mKRB supplemented with 10 to 20% sheep serum survived 48 hr in vitro, and they had more (P<.05) nuclei than those cultured in 0 or 50% sheep serum. mKRB containing 10% sheep serum and mKRB containing 10% fetal calf serum supported similar in vitro development; however, few blastocysts cultured in medium containing 10% pig serum survived. Swine morulae benefited when cultured in the presence of 10% sheep serum, as evidenced by an increase in the number of embryos hatching during culture (0 and 18.2% for mKRB and mKRB plus 10% sheep serum, respectively). Culturing day 10 and day 12 blastocysts in mKRB plus 10% sheep serum, however, was not so successful. Day 10 blastocysts recovered from one of two donor gilts increased in size and nuclei number in vitro, but none of the day 12 blastocysts survived 24 hr in vitro. These studies indicate that the requiremnts for swine embryo development in vitro change at about the time of blastocoel formation, since mKRB will not support development of day 8 blastocysts unless supplemented with serum, and that hatching percentage is improved by including serum in the culture medium for morulae. During day 10 to 11, there is another qualitative change in requirements of swine blastocysts such that mKRB plus serum will no longer support development.

¹ Contribution No. 80-342-j, Dept. of Anim. Sci. and Ind., Kansas Agr. Exp. Sta., Manhattan 66506.

² Present address: Dept. of Anim. Sci., Univ. of Illinois, Urbana 61801.

³ Dept. of Anim. Sci. and Ind.

⁴ Fraction V.