Pars Journal of Medical Sciences

Effect of Vero cell Co – culture on Quality of in vitro produced bovine blastocysts after vitrification

Authors

Abstract

Introduction : Post freze – thaw embryo quality can be assessed by various methods , the most common being in vetro survival after a period of culture . several culture system have been used for this purpose and most of them include somatic cell such as oviduct , granulosa and  Buffalo rat liver cells as Co- culture support . The aim of this study was to assess the Vero cell Co- culture system on quality of in vitro produced bovine blastocysts after vitrification.


Materials & Methods :after in vitro maturation and fertilization of Immature bovine oocytes . presumptive zygotes were cultured in SoFand SoF medium for 7 days (39 c , 5% Co) . Then 100 blastorcysts were selected and vitrified  ( 40% ethylene glycol , 18% ficol , 0.3 mol sacurose).After tawing blastocysts were divided into two groups . One group was cultured in SOF : and the other group was cultured in SoF + Vero cell medium for 48 h. re-expanded blastocysts were selected, inner cell mass(ICM) from them were recovered by immunosurgery and stained cell with Hoechst .


Results : The mean number of ICM in SoF and SoF + Vero were 12 and 15 respectively . The percentage re-expanded blastocysts in SoFand SoF+ Vero were 42%,54% respectivety . there were significant difference between the number of ICM and percentage re–expanded blastocysts in the two groups.


Conclusion : Vero cell Co – culture system can improve viability of in vitro produced bovine blastosysts after vitrification . 

Keywords

1- Wheeler, MB, Walters EM.Transgenic Technology and Application in swine . Theriogenology 2001; 56: 1345-1369. 2- Bleck GT,White BR, Miller DJ , wheeler MB. Production & Lactalbumin in the milk of transgenic pigs. J Anim sci 1998 ; 76 : 3072 -3078 . 3- Brenig B, Mullar M, Brem G. Gene transferin pigs proc 4th world cong genet Apptied to live stock prod , 1990 ; 12: 41-48. 4- Nguyen BX,sotomaruy , Tani T , Katoy Tsunoday . Efficient cryopreservation of bovine blastocysts drived form nuclear transfer with somatic cells using partial dehydration and vetrification . Theriogenology 2000 ; 53 : 1439 -1448. 5- Hovatta O,sily R,Kraus Z T.and et al . Gryopreservation of human ovarian tissue using dimethyl sulphoxid and propnediol socrose as cryoprotectants . Human Repord 1996 ; 11(6) : 1268-1272 . 6- Leibo SP. Field trial one step frozen bovine embryos transferred non-surgically . Theriogenology 1983;19:139. 7- Rizos D, ward F, Boland MP,Lanergand P.Effect of culture system on the yield and guality of bovine blastocysts as assessed by survival after vitrificaiton . Theriogenology 2001 ; 56:1-16. 8- Fahy GM. Cryopotectant toxicity . Biochemical or smatic ? cryo – lett . 1984;5:287-294. 9- Mtango NR,varisanga MD,Dong YJ , Rajamahendvan R Growth factors and growth hormon enhence in vitro embryo production and post than survival of vitrified bovine blastocysts. Theriogenology 2003;5:1393-1402 . 10- Parrish JJ , susko- parrish JL.first NL.Capacitition of bovin sperm by heparin .Biol Reprod . 1988 ; 138:1171-80. 11- Steeves TE,Gardner DK. Temporal and differential effects of amino acids on bovine embryo development in cultrure . Biol Reprod 1999 :61 : 731- 40 . 12- Zhu SE,Kasai M,otage H. Cryopreservation of expanded mouse blastocysts by vetrification in ethylenglycol – based solution.J Reprod Fertil 1993 ;98:139 -145. 13- Gutierrez .Adan lenergand P.Rizos D.Effect of the in vitro culture system on the kinetus of blastocyst development . Theriogenelogy 2001;55:1117 -1126. 14- Dobrinsky JR.Cellular approach to cryopreservation of embryos.Theriogenology 1996;45:17-26. 15- Shaha.S,suzuk, T.Vetrification of in vitro procluced bovine embryos at different ages using one and three – stop addition of cryoprotective additives.Reprod Fertil Dev 1997 ;9:741- 746. 16- Martinez AG,Valcarcel A,Valcarcel A,Brogliatti G.Vitrification of in vitro production bovine embryos in vitro and in vivo evaluation .Anim Reprod sci 2002 , 73:11-21. 17- Kaidi S,Donnay I,Vanlangendanck A,Massip A.camparison of two co- culture system to assess the survival of in vitro produced bovine blastocysts after vitrification . Anim Reprod sci 1998;52:39-50. 18- Vajta G, Hom P,Grev T. Overall efficiency of in vitro embryo production and vitrification in cattle . Theriogenology 1996;45:683-689. 19- Hiroyoshi H, In vitro production of bovine embryos and their application for embryo transfer. Theriogenology 2003 59:675-685.