Zakiyatul Faizah, Ninik Darsini, Aucky Hinting

= http://dx.doi.org/10.20473/fmi.v52i2.5222
Abstract views = 561 times | downloads = 506 times


The success rate of fertilization post save frozen oocytes is still very low, because the oocyte has distinctive features, namely the volume ratio and a lower surface to the limited penetration of water and cryoprotectants penetrate cells. Beside mature oocytes have a thread spindles are particularly vulnerable to the drop in temperature. Keep frozen oocytes is needed, especially in women who needed rescue fertility so their oosit can be fertilized. Maturation is done in TC 100 mL medium covered with mineral oil in a petri dish with a diameter of 36 mm. Oocyte vitrification begins with washing in PBS supplemented medium serum 20% for 1-2 minutes, followed by serum in the medium PBS + 20% + 10% ethylene glycol for 10-14 minutes. Then oocyte vitrification medium is transported in PBS + serum 20% + sucrose 0.5M ethylene glycol + 15% + 15% PROH for 25-30 seconds. Thawing oocytes is done by successive immersed in the media: 1). PBS + 20% serum + 0.5M sucrose, 2). PBS + 20% serum + 0.25M sucrose, and 3). PBS + 20% serum + 0.1 M sucrose. Insemination is done in rosset, and the number of fertilization was observed after 48 hours. Fertilization in the control group amounted to 42.97%, while the K1 and K2 there are no fertilization at all. The analysis showed that fertilization in the control and treatment groups significantly different at p <0.05 in both treatment groups K1 or K2 there are no fertilization at all. The conclusions of this study is there is no difference between the amount of fertilization of bovine oocytes were vitrified pre and post-maturation in vitro.


vitrified, In vitro fertilization

Full Text:



Carrel TD and Peterson CM (2010). Reproductive Endocrinology and Infertility, New York, Springer, p 635

Chang CC, Lin CJ, Sung LY, Kort HI, Tian XC, Nagy ZP (2011). Impact of phase transition on the mouse oocyte spindle during vitrification. Reproductive Biomedicine Online 22, 184-191

Chen SU, Lien YR, Chao KH, Ho HN, Yang YS, Lee TY (2003). Effects of cryopreservation on meiotic spindles of oocytes and its dynamics after thawing: clinical implications in oocyte freezing–a review article. Molecular and Cellular Endocrinology 202, 101–107

Coticchio G, Bonu MA, Borini A, Flamigni C (2004). Oocyte cryopreservation: a biological perspective. European Journal of Obstetrics & Gynecology and Reproductive Biology 115, S2-S7

Fujiwara K, Seita Y, Inomata T, Junya ITO, Kashiwazaki N (2010). Ethylene glycol-supplemented calcium-free media improve zona penetration of vitrified rat oocytes by sperm cells. Journal of Reproduction and Development 56, 169-175

Ghetler Y, Yavin S, Shalgi R, Arav A (2005). The effect of chilling on membrane lipid phase transition in human oocytes and zygotes. Human Reproduction 20, 3385–3389

Ghetler Y, Skutelsky E, Nun IB, Dor LB, Amihai D, Shalgi R (2006). Human oocyte cryopreservation and the fate of cortical granules. Fertility and Sterility 86, 210-216

Hu W, Marchesi D, Qiao J, Feng HL (2012). Effect of slow freeze versus vitrification on the oocyte: an animal model. Fertility and Sterility 98, 752-760

Kuwayama M, Vajta G, Kato O, Leiobo SP (2005). Highly efficient vitrification method for cryopreservation of human oocyte. Reproductive Biomedicine Online 11, 300-308

Kohaya N, Fujiwara K, Junya ITO, Kashiwazaki N (2011). High developmental rates of mouse oocytes cryopreserved by an optimized vitrification protocol: the effects of cryoprotectants, calcium and cumulus cells. Journal of Reproduction and Developmet 57, 675-680

Larman MG, Katz-Jaffe MG, Sheehan CB, Gardner DK (2007). 1,2-propanediol and the type of cryopreservation procedure adversely affect mouse oocyte physiology. Human Reproduction 22, 250-259

Larman MG, Sheehan CB, Gardner DK (2006). Calcium-free vitrification reduce cryoprotectant-induced zona pellucida hardening and increases fertilization rates in mouse oocytes. Reproduction 131, 53-61

Mavrides A and Morroll D (2005). Bypassing the effect of zona pellucida changes on embryo formation following cryopreservation of bovine oocytes. European Journal of Obstetric & Gynecology and Reproductive Biology 118, 66-70

Men H, Monson RL, Parrish JJ, Rutledge JJ (2003). Degeneration of cryopreserved bovine oocytes via apoptosis during subsequent culture. Cryobiology 47, 73-81

Niwa K, Oghada O, Yuhara M (1988). Effects of caffeine in media for pretreatment of frozen-thawed sperm on in-vitro penetration of cattle oocytes. Proc 11th Int Congr Anim Reprd Al 3, 346

Parrish JJ, Krogenaes A, Susko-Parrish JL (1995). Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development. Theriogenology 44, 859-869

Prokofiev MI, Ernst LK, Suraeva NM, Lagutina IS, Udavlennikova NN, Kesyan AZ, Dolgohatskiy AI (1992). Bovine oocyte maturation, fertilization and further development in vitro and after transfer into recipients. Theriogenology 38, 461-469

Tian SJ, Yan CL, Yang HX, Zhou GB, Yang ZQ, Zhu SE (2007). Vitrification solution containing DMSO and EG can induce parthenogenetic activation of in vitro matured ovine oocytes and decrease sperm penetration. Animal Reproduction Science 101, 365-371

VandeVoort CA, Shirley CR, Hill DL, Leibo SP (2007). Effects of cryoprotectants and cryopreservation on germinal vesicle-stage cumulus–oocyte complexes of rhesus monkeys. Fertility and Sterility 90, 805-816


  • There are currently no refbacks.

Copyright (c) 2017 Zakiyatul Faizah, Ninik Darsini, Aucky Hinting

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.

Indexed By

View My Stats

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.