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United States Department of Agriculture

Agricultural Research Service

Title: Production of Piglets Preselected for Sex Following in Vitro Fertilization with Flow Cytometrically Sorted X and Y Chromosome Bearing Sperm

item Johnson, Lawrence
item Dobrinsky, John
item Welch, Glenn
item Niemann, H - MARIENSEE, GERMANY

Submitted to: Theriogenology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: October 24, 1996
Publication Date: N/A

Interpretive Summary: Gender preselection in swine is a useful technology for producing male and female lines of hybrid stock. Additionally, the female provides a faster growth rate and feed efficiency. The combination of the sexing technology and in vitro fertilization was shown to be an effective means of producing litters of one sex only. Two litters totaling 10 females were born. These elitters were the first produced from sexed embryos based on the Beltsville Sexing Technology. These results will be used by scientists to further perfect the combination of in vitro fertilization in the pig with our current sexing technology.

Technical Abstract: In vivo matured porcine oocytes were fertilized in vitro with X and Y chromosome bearing sperm, flow cytometrically sorted for sex based on DNA content. Developmental competence of the sexed embryos was determined through established pregnancies after embryo transfer. Sperm were stained with Hoechst 33342 and sorted using a flow cytometry cell sorter. Purity of sort was 83% for Y-sperm and 92% for X-sperm, respectively. A total of 387 mature cumulus-oocyte-complexes were collected from 18 superovulated prepuberal gilts shortly before ovulation. In vitro fertilization with sorted sperm was performed in four replicates. After 18 h of sperm and egg co-culture at 39 degrees C, zygotes were placed into culture medium (NCSU- 23) for anther 24 h. The average cleavage rate was 56.2%. Ninety-two embryos produced from X-sorted sperm were transferred surgically into the uterus of 2 recipients. Two gilts farrowed and delivered 6 and 4 healthy female piglets, respectively. Additionally, two gilts were inseminated intratubally via surgical laparotomy with either X or Y sorted sperm (2x100,000) per oviduct. The two sows farrowed and litters were born producing 15 piglets. Thirteen of the 15 piglets were of the predicted gender (85%).

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