Isachenko, V.; Lapidus, I.; Isachenko, E.; Hancke, K.; Nawroth, F.; Wiegratz, I.; Kreienberg, R.
October 2010
Reproductive BioMedicine Online (Reproductive Healthcare Limited;Oct2010 S3 Supplemen, Vol. 20, pS54
Academic Journal
Objective: Cryopreservation, the most important stage of the cryobanking of ovarian tissue, can be carried out using one of two methods: conventional ("slow") freezing, and vitrification (direct plunging into liquid nitrogen). For human oocytes vitrification is more effective compared to conventional freezing (Isachenko et al., 2005a,b; 2006a). However, these comparative data are limited for human ovarian tissue. It was previously shown that for human ovarian tissue the effectiveness of conventional freezing is higher compared to vitrification (Isachenko et al., 2007, 2009a, b). However, this question (what is better, traditional freezing or modern vitrification?) is topical and at present the respective discussions in the field of reproductive medicine are more than intensive. Particularly, the results of our comparative investigations (freezing vs vitrification) (Isachenko et al., 2009a) were agitated a criticism of some colleges, by opinion of which it is not correct to compare the protocol of vitrification developed by us (Isachenko et al., 2008) with standard conventional freezing. By opinion of some colleges, there are effective protocols of vitrification of human ovarian tissue. Taking into account this critical opinion, the aim of our investigations was the comparison of effectiveness of recently published and, by analyse of literature, most effective protocol of vitrification of human ovarian tissue (Keros et al., 2009c, traditionally experienced group from Sweden) and conventional freezing with our modifications (Isachenko et al., 2009a). Materials and Methods: Ovarian tissue from 5 patients was transported to the laboratory within 20 min at 32 to 34°C, divided into smaller pieces (1x1 to 1.5x0.7 to 1mm) and randomly distributed into three groups: Group 1: control (fresh tissue), Group 2: pieces after vitrification/warming, Group 3: pieces after conventional freezing/thawing. All pieces were cultured in vitro for 12 days (Isachenko et al., 2006b). The viability of the tissue was evaluated by the development of the follicles and GAPDH gene expression after in vitro culture. Total RNA preparation, RT, PCR, and real-time PCR were performed by the routine methodology using GenElute Total RNA purification-, DNase I treatment-, Superscript II reverse transcriptase-, PCR RedTaq Ready Master Mix- and SYBR Master Mix- Kits.


Related Articles

  • Mouse Ovarian Tissue Cryopreservation Has Only a Minor Effect on In Vitro Follicular Maturation and Gene Expression. Hung-Ching Liu; Zhiming He, Larry I.; Rosenwaks, Zev // Journal of Assisted Reproduction & Genetics;Oct2003, Vol. 20 Issue 10, p421 

    Purpose: To establish a protocol for ovarian tissue cryopreservation which can retain fertility potential after thawing and to evaluate the impact of cryopreservation on development and gene expression during folliculogenesis. Methods: A controlled randomized study in a clinical and academic...

  • Agentes crioprotetores intracelulares: características e utilização na criopreservação de tecido ovariano e oócitos. Castro, Simone Vieira; de Andrade Carvalho, Adeline; Gomes da Silva, Cleidson Manoel; Rocha Faustino, Luciana; de Figueiredo, José Ricardo; Ribeiro Rodrigues, Ana Paula // Acta Scientiae Veterinariae;Mar2011, Vol. 39 Issue 2, Special section p1 

    Background: The application of cryopreservation in human and animal reproductive medicine has stimulated several studies about the effects of low temperatures and freezing processes on cells and tissues, in order to develop efficient protocols for gamete and embryo preservation. Moreover,...

  • Oocyte cryopreservation: the birth of the first Hungarian babies from frozen oocytes. Konc, Janos; Kanyo, Katalin; Varga, Erika; Kriston, Rita; Cseh, Sandor // Journal of Assisted Reproduction & Genetics;Jul2008, Vol. 25 Issue 7, p349 

    Purpose To present data obtained with clinical application of oocyte cryopreservation. Methods Slow freezing/rapid thawing in PBS based medium containing 1.5 M propanediol+0.3 M sucrose. Results A total of 127 embryos were transferred into 54 patients (1.9 embryo/cycle, 64 transfer cycles)....

  • Evaluation of MMP-2 gene expression in mice pre-antral follicles derived from vitrified and fresh ovarian tissue. Khosravi, S.; Zavareh, S.; Paylakhi, S.; Gorbanian, M. // Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p84 

    Introduction: Cryopreservation is a useful method for preservation of ovarian tissue of patients who are candidate for chemo and radiotherapy. It has been postulated that MMP-2 participates in the process of remodeling of ovarian extracellular matrix such as the follicle growth and ovulation....

  • Human ovarian tissue cryopreservation: quality of follicles as a criteria of effectiveness. Isachenko, V.; Isachenko, E.; Kreienberg, R.; Woriedh, M.; Weiss, J. M. // Reproductive BioMedicine Online (Reproductive Healthcare Limited;Apr2010, Vol. 20 Issue 4, p441 

    Experiments comparing vitrification and conventional freezing of mammalian ovarian tissue show that vitrification can also guarantee the storage of viable follicles after warming, but conventional freezing is more effective. The central goat of cryotechnology is the preservation of intact...

  • Cryopreservation of intact human ovary with its vascular pedicle. Mohamed A. Bedaiwy; Mahmoud R. Hussein; Charles Biscotti; Tommaso Falcone // Human Reproduction;Dec2006, Vol. 21 Issue 12, p3258 

    BACKGROUND: The aim of this study was to assess the immediate post-thawing injury to the human ovary that was cryopreserved either as a whole with its vascular pedicle or as ovarian cortical strips. MATERIALS AND METHODS: Bilateral oophorectomy was performed in two women (46 and 44 years old)...

  • Vitrification as an alternative means of cryopreserving ovarian tissue. Amorim, Christiani A; Curaba, Mara; Van Langendonckt, Anne; Dolmans, Marie-Madeleine; Donnez, Jacques // Reproductive BioMedicine Online (Reproductive Healthcare Limited;Aug2011, Vol. 23 Issue 2, p160 

    Because of the simplicity of vitrification, many authors have investigated it as an alternative to slow freezing for cryopreserving ovarian tissue. In the last decade, numerous studies have evaluated vitrification of ovarian tissue from both humans and animals. Different vitrification solutions...

  • Five-Year Experience with a Sodium-Depleted Slow-Freezing Method.  // Fertility Weekly;10/30/2006, p7 

    The article presents a study that examined the effectiveness of oocyte cryopreservation by the use of sodium-depleted freezing media. Data used came from 53 frozen egg-embryo transfer cycles in 46 patients that were carried from April 2000 to August 2005. Patients underwent fresh non-donor...

  • Mammalian Oocyte Cryopreservation - Review. Cean, Ada; Bogdan, Alexandru T.; Pacala, Nicolae; Ivan, Alexandra; Ilie, Daniela E. // Scientific Papers: Animal Science & Biotechnologies / Lucrari St;Jun2011, Vol. 44 Issue 1, p370 

    Although oocyte cryopreservation represents one of the main objectives of the reproductive medicine and cryobiology in the last years, until now, regardless of the studied specie, there is no freezing protocol that will assure satisfactory survival rates after thawing. Oocyte cryopreservation is...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics