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2012 | 68 | 01 |

Tytuł artykułu

Próba uzyskania zarodków końskich po zapłodnieniu wspomaganym (ICSI)

Warianty tytułu

EN
Attempt to obtain equine embryos after assisted fertilization (ICSI)

Języki publikacji

PL

Abstrakty

EN
The aim of this study was to obtain embryos after assisted fertilization of oocytes collected from slaughtered mares and matured in vitro by an intracytoplasmic sperm injection (ICSI) and to establish optimal conditions for the in vitro culture of the embryos. Oocytes were collected by the scraping method and cultured in vitro in TCM with Earl’s salts supplemented with 20% FBS, 5 g/ml FSH, estradiol 1 g/ml, pyruvate and glutamine. After a 30-hour in vitro culture oocytes that had reached the metaphase II stage were subjected to an intracytoplasmic sperm microinjection (ICSI) with frozen semen. After thawing, a motile sperm selection was carried out by the „swim up” method. Then the sperm was immobilized, aspirated into an injection pipette, and introduced into the cytoplasm of an oocyte. Microinjected oocytes were transferred onto a culture medium: DMEM F12 or SOF. The control group consisted of oocytes subjected to microinjection without sperm (sham ICSI). Oocytes that did not show signs of development and embryos in different developmental stages were fixed and stained with Hoechst fluorochrome. The in vitro culture of embryos in the DMEM F12 medium produced only early stages of development (pronucleus, two-blastomere embryos). In contrast, the in vitro culture in the SOF medium produced embryos in the morula stage (6.8%). In the control group, no signs of embryonic development were observed.

Wydawca

-

Rocznik

Tom

68

Numer

01

Opis fizyczny

s.54-57,fot.,tab.,bibliogr.

Twórcy

autor
  • Katedra Rozrodu i Anatomii Zwierząt, Wydział Hodowli i Biologii Zwierząt, Uniwersytet Rolniczy w Krakowie, Al.Mickiewicza 24/28, 30-059 Kraków
autor
autor
autor

Bibliografia

  • 1.Battut I., Bézard J., Palmer E.: Establishment of equine oviduct cell monolayers for co-culture with early equine embryos. J. Reprod. Fertil. Suppl. 1991, 44, 393-403.
  • 2.Choi Y. H., Love C. C., Love L. B., Varner D. D., Brinsko S., Hinrichs K.: Developmental competence in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection with fresh or frozenthawed spermatozoa. Reproduction 2002, 455-465.
  • 3.Choi Y. H., Love C. C., Varner D. D., Hinrichs K.: Equine blastocyst development after intracytoplasmic injection of sperm subjected to two freeze-thaw cycles. Theriogenology 2006, 65, 808-819.
  • 4.Choi Y. H., Love C. C., Varner D. D., Love L., Hinrichs K.: Effects of gas conditions, time of medium change, and ratio of medium to embryo on in vitro development of horse oocytes fertilized by intracytoplasmic sperm injection. Theriogenology 2003, 59, 1219-1229.
  • 5.Choi Y. H., Roasa L. M., Love C. C., Varner D. D., Brinsko S. P., Hinrichs K.: Blastocyst formation rates in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection. Biol. Reprod. 2004, 70, 1231-1238.
  • 6.Dell'Aquila M. E., Cho Y. S., Minoia P., Traina V., Lacalandra G. M., Maritato F.: Effects of follicular fluid supplementation of in vitro maturation medium on the fertilisation and development of equine oocytem after in-vitro fertilization and intracytoplasmic sperm injection. Hum. Reprod. 1997, 12, 2766-2772.
  • 7.Franz L. C., Choi Y. H., Squires E. L., Seidel G. E., Hinrichs K.: Effects of roscovitine on maintenance of the germinal vesicle in horse oocytes, subsequent nuclear maturation, and cleavage rates after intracytoplasmic sperm injection. Reproduction 2003, 125, 693-700.
  • 8.Galli C., Crotti G., Turini P., Duchi R., Mari G., Zavaglia G., Duchami G., Deels P., Lazzari G.: Frozen-thawed embryos produced by ovum pick up of immature oocytes and ICSI are capable to establish pregnancy in the horse. Theriogenology 2002, 58, 705 (Abstr).
  • 9.Galli C., Lazzari G.: In vitro and in vivo culture in the sheep oviduct of equine embryos obtained by IVM and ICSI. 2nd Meeting Europ. Equine Gamete Group: 26th-29th Sept. 2001. Loosdrecht, The Netherlands. Abstr. s. 55-56.
  • 10.Grondhal C., Hansen T., Hossanini A., Heinze I., Greve T., Hyttel P.: Intracytoplasmatic sperm injection of in vitro maturated equine oocytes. Biol. Reprod. 1997, 57, 1495-1501.
  • 11.Guignot F., Ottogalli M., Yvon J. M., Magistrini M.: Preliminary observations in vitro development of equine embryo transfer after ICSI. Reprod. Nutr. Dev. 1998, 38, 653-663.
  • 12.Hinrichs K., Choi Y. H., Love L. B., Varner D. D., Love C. C., Walckenaer B. E.: Chromatin configuration within the germinal vesicle of horse oocytes: changes post mortem and relationship to meiotic and developmental competence. Biol. Reprod. 2005, 72, 1142-1150.
  • 13.Hinrichs K., Love C. C., Brinsko S. P., Choi Y. H., Varner D. D.: In vitro fertilization of in vitro matured equine oocytes: Effect of Maturation medium, duration of maturation, and sperm calcium ionophore treatment, and comparison with rates of fertilization in vitro after oviductal transfer. Biol. Reprod. 2003, 67, 256-262.
  • 14.HiramotoY.: Microiniection of the live spermatozoa into sea urchin eggs. Exp. Cell. Res. 1962, 27, 416-426.
  • 15.Kimura Y., Yanagimachi R.: Development of normal mice from oocytes injected with secondary spermatocyte nuclei. Biol. Reprod. 1995, 53, 855-862.
  • 16.Lazzari G., Grotti G., Turini P., Duchi R., Mari G., Zavaglia G., et.al.: Equine embryos at the compacted morula and blastocyst stage can be obtained by intracytoplasmic sperm injection (ICSI) of in vitro matured oocytes with frozen-thawed spermatozoa from semen of different fertilities. Theriogenology 2002, 58, 709-712.
  • 17.Li X., Morris L. H. A., Allen W. R.: Influence of co-culture during maturation on the developmental potential of equine oocytes fertilised intracytoplasmic sperm injection (ICSI). Reproduction 2001, 121, 925-932.
  • 18.Palermo G., Joris H., Devroey P., Van Steirteghem A. C.: Pregnancies after intracytoplasmic sperm injectionof single spermatozoon into a oocyte. Lancet 1992, 340, 17-18.
  • 19.Rosati I., Berlinguer F., Bogliolo L., Ledda S., Naitana S.: The effect of co-culture on the development of in vitro matured equine oocytes after intracytoplastic sperm injection. Equine Vet. J. 2002, 34, 73-78.
  • 20.Squires E. L., Wilson J. M., Kato H.: A pregnancy after intracytoplasmic sperm injection into equine oocytes matured in vitro. Theriogenology 1996, 45, 306 (Abstr).

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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