Histological description of aceh cattle ovary cryopreserved by various cryoprotectants

DOI: 10.18805/ijar.B-688    | Article Id: B-688 | Page : 1223-1226
Citation :- Histological description of aceh cattle ovary cryopreserved by various cryoprotectants.Indian Journal Of Animal Research.2018.(52):1223-1226
Syafruddin Syafruddin, Tongku Nizwan Siregar, Azrina Azrina, Teuku Armansyah, Budianto Panjaitan, Dwinna Aliza , Amalia Sutriana, Zuhrawati Zuhrawati, Rosmaidar Rosmaidar, Roslizawaty Roslizawaty roslizakh@unsyiah.ac.id
Address : Laboratory of Clinic, Veterinary Medicine Faculty, Syiah Kuala University, Banda Aceh, Indonesia.
Submitted Date : 7-01-2017
Accepted Date : 19-02-2018

Abstract

This research aimed to determine Aceh cattle ovarian follicle morphological integrity after vitrified by various cryoprotectants. Cryoprotectants used in this research were 30% ethylene glycol (EG), 30% dimethyl suphocide (DMSO), and combination of 15% EG + 15% DMSO. Prior to vitrification process, ovaries were cleansed by phosphate buffered saline (PBS) and then cut into ±1 mm³ . Ovaries were consecutively submerged into the following liquid for 5 minutes each: PBS+ 0.25 M sucrose; PBS+ 0.5 M sucrose; PBS+ 0.5 M sucrose + 10% cryoprotectants; and PBS+ 0.5 M sucrose + 30% cryoprotectants. Then, ovaries were packed into straws with ±7 cm in length and ± 6 mm in diameter. Before kept in liquid nitrogen, ovaries were first exposed to nitrogen fume for 10 second. After being stored for 1 day, the ovaries were proceed for histological examination . The result showed that Aceh cattle ovarian follicle after vitrification using 30% EG  yields the best morphological integrity. Cumulus oophorus, zona pellucida, granulose cell arrangement, theca interna, and theca externa cells were observed clearer in ovary which was vitrified with 30 % EG than those with 30% DMSO  and combination of 15% EG + 15% DMSO. As conclusion, 30% EG  was able to protect ovary morphological integrity better than 15 % EG + 15% DMSO  and 30% DMSO. Furthermore, combination of 15% EG+ 15 % DMSO  was relatively better in protecting ovary follicle morphological integrity compared to 30% DMSO.

Keywords

Aceh cattle Cryoprotectant Ovary Vitrification.

References

  1. Dattena M, Accardo C, Pilichi S. (2004). Comparison of different vitrification protocols on viability after transfer of ovine blastocysts in vitro produced and in vivo derived. Theriogenology. 62: 481-493.
  2. Djuwita I, Boediono A, Sukra Y.(2000). Morphology and fertilization rate of vitrified ovine oocytes matured in vitro. 14th International Congress on Animal Reproduction, Stockholm. 2: 202-207.
  3. Donnez J, Dolmans MM, Squifflet J. (2006). Restoration of ovarian function after orthotopic (intraovarian and priovarian) transplantationof cryopreserved ovarian tissue in a women treated by bone marrow transplantation for sickle cell anemia. Hum Reprod. 21: 183-188.
  4. Fukui EJ, Xia L, Downey BR. (1993). Ultrastructural changes in bovine oocyte cryopreserved by vitrification. Cryobiology. 32: 139-    156.
  5. Gao D, Critser JK. (2000). Mechanisms of cryoinjury in living cells. ILARJ. 41: 187-196.
  6. Ghorbani M, Sadrkhanlou R, Nejati V, Ahmadi A, Tizroo G. (2012). The effects of dimethyl sulfoxide and ethylene glycol as vitrification protectants on different cleavage stages of mouse embryo quality. Vet Res Forum. 3: 245-249.
  7. Hyttel P, Fair T, Greve T. (1997). Oocyte growth, capacitation and final maturation in cattle. Theriogenology. 47: 23-32.
  8. Jain JK, Paulson RJ. (2006). Oocyte cryopreservation. Fertil Steril. 86: 1037-1046.
  9. Kasai M. (2002). Advances in the cryopreservation of mammalian oocytes and embryos: Development of ultra rapidvitrification. Reprod Med Biol. 1: 1-9.
  10. Lucci CM, Kacinskis MA, Rumpf R. (2004). Effect of different cryoprotectants on the structural preservation of follicles in frozen zebu bovine (Bosindicus) ovarian tissue. Theriogenology. 61: 1101-1114.
  11. Lucci CM, Schreier LL, Machado GM. (2007) Effects of storing pig ovaries at 4 or 20°C for different periods of time on the morphology and viability of pre-antral follicles. Reprod Dom Anim. 42: 76-82.
  12. Mohamad K, Rumiyati E, Sari F. (2000). Kriopreservasi Oosit pronukleus dan embrio mencit dengan metode vitrifikasi. Seminar Nasional Biologi XVI Bandung. 45-49.
  13. Mohamad K, Djuwita I, Boediono A. (2005). Vitrifikasi ovarium mencit menggunakan etilenglikol dan DMSO sebagai krioprotektan dan viabilitasnya pasca auto transplantasi di subkapsulaginjal. Med Ked Hewan. 21: 23-27.
  14. Rosadi B, Setiadi MA, Sajuthi D. (2011). Presevasi ovarium dan pengaruhnya terhadap morfologi folikel domba. J. Vet. 12: 11-15.
  15. Takeda T. (1990). Preparing Cryoprotectants. In Techniques for Freezing Mammalian Embryos. Seidel, G.E. (Ed.). Colorado State University, USA. 97-102.
  16. Wahjuningsih S, Harjopranjoto S, Sumitro SB. (2010). Pengaruh konsentrasi etilen glikol dan lama paparan terhadap tingkat fertilitas in vitro oosit sapi. J Ked Hewan. 4: 12-17.
  17. Vajta G , Im PH, Kuwayama M. (1998). Open pulled straw (OPS) vitrification: A new way to reduce cryo injuries of bovine ova and embryos. Mol Reprod Dev. 51: 53-58. 

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