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Coculture of Bovine Demiembryos Prior to Freezing
»Frysning af halve kvægembryoner efter forudgående dyrkning«.
Acta Veterinaria Scandinavica volume 33, pages 237–243 (1992)
Abstract
Pregnancy rates after transfer of frozen/thawed bisected embryos, demiembryos, have until now been very low. In the present study it was attempted to improve the freezability rate of demiembryos by culturing them on a monolayer of bovine oviduct epithelial cells (BOEC) prior to freezing. The cultured frozen/thawed demiembryos showed a lower developmental rate than intact embryos. The pregnancy rate (23% ) following transfer was not different from the pregnancy rate after transfer of unfrozen demiembryos (26% ). The calving rate (4%), however, was significantly lower than the calving rate after transfer of fresh demiembryos (23%). Although the overall pregnancy rate achieved in this study was low, it can be concluded that a co-culture period upon BOEC prior to freezing does not improve the viability of frozen demiembryos.
Sammendrag
Halve embryoner, demiembryoner, som fryses, optøs og overføres, giver en meget lav drægtighedsprocent. Formålet med dette forsøg var at undersøge, om det var muligt at forbedre frysbarheden af demiembryoner ved at dyrke dem i 24 t med bovine Ovidukt epithel celler (BOEC). Dette dyrkningssystem skulle give demiembryonerne mulighed for at regenerere og i løbet af dyrkningsperioden forøge celleantallet. På denne måde skulle de gøres mere egnede til at overleve fryseskaderne. Deres viabilitet efter frysning blev vurderet dels ved drægtigheds procent efter overførsel og dels ved udviklingsevne under 24 t genoptaget dyrkning. Ved dyrkning af 243 demiembryoner viste det sig, at de både før og efter frysning havde en dårligere udviklingkapacitet end intakte embryoner.
Ved overførsel af 48 dyrkede og frosne demiembryoner opnåedes en drægtighedsfrekvens på 23%, mens kælvingsfrekvensen kun var på 4%. Ved overførsel af 142 friske demiembryoner var de tilsvarende tal henholdsvis 26% og 23%. Det kunne tolkes således, at demiembryonerne efter dyrkning og frysning har celler nok til at danne fosterblære, men for få til at danne selve fosteret.
Forsøget viste således, at dyrkning med BOEC ikke forbedrede frysbarheden af halve bovine embryoner.
References
Albihn A, Rodriguez-Martinez H, Gustafson H: Morphology of day 7 bovine demi-embryos during in vitro reorganization. Acta anat. 1990, 138, 42–49.
Albihn A, Gustafsson H, Rodriguez-Martinez H: Morphology of day-7 bovine demi-embryos and their post-transfer Viability. Reprod.dom. Anim. 1991, 26, 85–87.
Bielanski A, Hare W C D: Survival in vitro of bovine demiembryos after freezing by slow cooling rates or vitrification. Theriogenology 1988, 29, 223.
Chesne P, Heyman Y, Chupin D, Procureur R, Menezo Y: Freezing of cattle demiembryos: Influence of a period of culture betwween splitting and freezing on survival. Theriogenology, 1987, 27, 218.
Heyman Y, Chesne P: Development of cattle embryos after splitting or throphoblast biopsy and freezing. 11th Int. Congr. Anim. Repr. and A.I., Dublin, 1988, 2, 167.
Holm P, Greve T, Bak A, Schmidt M: Bisection of bovine morulae and blastocysts from superovulated danish dairy cows. Acta vet. scand. 1991, 32, 47–53.
Lehn-Jensen H, Willadsen S: Deep-freezing of cow ‘-half and ‘quarter’ embryos. Theriogenology, 1983, 19, 49–54.
Lehn-Jensen H: Cryopreservation of bovine embryos The Royal Veterinary and Agricultural University Copenhagen. Thesis. 1986.183 pp.
McEvoy T G, Sreenan J M: Effect of embryo quality and stage of development on the survival of zona pellucida-free cattle demi-embryos. Theriogenology, 1990, 33, 1245–1253.
Nieman H, Brem G, Sacher B, Smidt D, Krausslich H: An approach to sussecfull freezing of demiembryos derived from day-7 bovine embryos. Theriogenology, 1986, 25, 519–524.
Picard L, King A, Betteridge K.J.: Production of sexed calves from frozen/thawed embryos. Vet. Rec. 1985, 117, 603–608.
Picard L, Schneider U, Betteridge K J, King W A : Effect of the zona pellucida, agar embedding and culture on the survival of micromanipulated bovine embryos after freezing and thawing. J. In Vitro Fertilization and Embryo Transfer, 1988, 5, 268–274.
Schmidt M, Avery B, Purwantara B, Roschlau K, Greve T: In vitro culture of bovine demi-embryos on bovine oviduct ephithelial feeder cells. Reprod.dom.Anim. 1990, 25, 167–172.
Schmidt M, Avery B, Smith S: Culture of frozen/-thawed bovine demi-embryos with bovine oviduct epithelial cells. Reprod. dorn. Anim. 1991, 26, 88–91.
Schröder A, Miller J R, Thomsen P D, Roschlau K, Avery B, Poulsen P H, Schmidt M, Schwerin M.Sex determination of bovine embryos using the polymerase chain reaction. Anim. Biotech. 1990, 1, 121–133.
Schrøder U.: Cryopreservation of intact and splitted bovine embryos with glycerin and Gelee Royal. Inaug.-Diss. Hannover, 1988.
Schwidersky H, Seifert F, Genz S: Bovine embryo splitting-International trend and own results. Arch. Tierz, Berlin, 1990, 33, 139–48.
Seike N, Sakai M, Kanagawa H: Development of frozen-thawed demiembryos and production of identical twin calves of different age. J.vet.med.Sci.1991, 53, 37–42.
Shelton, J N: Factors affecting viability of fresh and frozen/thawed sheep demi-embryos. Theriogenology, 1992, 37, 713–721.
Smith S D, Schmidt M, Greve T: Transfer of bisected cattle embryos within the same zona pellucida. Reprod.Dom.Anim. 1991, 26, 91–93.
Willadsen S: Micromanipulation of embryos of the large domestic species. Diss. The Royal Veterinay and Agricultural University, Copenhagen (1982)
SAS Institute Inc: SAS/STAT Guide for Personal Computers, Version 6.03. Cary, NC, USA, 1988.
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Schmidt, M., Smith, S.D., Avery, B. et al. Coculture of Bovine Demiembryos Prior to Freezing. Acta Vet Scand 33, 237–243 (1992). https://doi.org/10.1186/BF03547314
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DOI: https://doi.org/10.1186/BF03547314