Cytogenetic behavior of cryoprotectant DMSO

IVF (in vitro fertilization) is now used worldwide to overcome female or male infertility. Cryopreservation of human embryos provides the clearest opportunity to improve the clinical results obtained with IVF. Cryoprotective agents (CPA) are used to minimize freezing injuries. DMSO has been the most widely used CPA, however, high concentrations of CPAs in the vitrification solution have been shown to be detrimental to the cell. In order to determine the effect of DMSO solutions (5%, 10% and 20%) on genetic stability and/or subsequent DNA repair, we have investigated its ability to induce Sister Chromatid Exchanges (SCEs) and Proliferation Rate Index (PRI) in normal human lymphocyte cultures of peripheral blood, due to the fact that the study cannot be conducted on embryos and to the limited number of spare available embryos, the corresponding accessible experimental material was T lymphocyte. The blood samples were taken from three different healthy donors (conducting experimental procedure in triplicate). After the effect of DMSO solutions on blood according to the instructions of kit K-SIBV-500, lymphocytes are harvested and cultured with suitable technique to assess SCEs and PRI. The results show that all three DMSO concentrations cause a statistically dose depended significant increase of SCE frequency of the lymphocytes (p<0.001) and raise the need for more research regarding the safe and effective use of cryoprotectant.

[1]  J. Deprest,et al.  Safe and effective cryopreservation methods for long‐term storage of human‐amniotic‐fluid‐derived stem cells , 2015, Prenatal diagnosis.

[2]  N. Greer Freezing under pressure: a new method for cryopreservation. , 2015, Cryobiology.

[3]  G. Almahbobi,et al.  Optimising vitrification of human oocytes using multiple cryoprotectants and morphological and functional assessment. , 2013, Reproduction, fertility, and development.

[4]  Z. Iakovidou-Kritsi,et al.  A comparative study on the cytogenetic activity of three benzodiazepines in vitro. , 2011, Genetic testing and molecular biomarkers.

[5]  G. Fahy Cryoprotectant toxicity neutralization. , 2010, Cryobiology.

[6]  C. Aragona,et al.  Ongoing pregnancies after vitrification of human oocytes using a combined solution of ethylene glycol and dimethyl sulfoxide. , 2006, Fertility and Sterility.

[7]  A. Massip Cryopreservation of embryos of farm animals. , 2001, Reproduction in domestic animals = Zuchthygiene.

[8]  D. Branch,et al.  Hematopoietic progenitor cells are resistant to dimethyl sulfoxide toxicity , 1994, Transfusion.

[9]  G. Fahy,et al.  Cryoprotectant toxicity and cryoprotectant toxicity reduction: in search of molecular mechanisms. , 1990, Cryobiology.

[10]  L. Giudice,et al.  Cryopreservation of embryos and ova. , 1988, Fertility and sterility.

[11]  P. Quinn,et al.  Experience with the cryopreservation of human embryos using the mouse as a model to establish successful techniques , 1986, Journal of in Vitro Fertilization and Embryo Transfer.

[12]  L. Murdoch Dimethyl sulfoxide (DMSO)--an overview. , 1982, The Canadian journal of hospital pharmacy.

[13]  S. Maeda,et al.  Factors involved in differential giemsa-staining of sister chromatids , 1978, Chromosoma.

[14]  B. Tarlatzis,et al.  Cytoskeletal analysis of human blastocysts by confocal laser scanning microscopy following vitrification. , 2012, Human reproduction.

[15]  G. Vajta,et al.  Vitrification in assisted reproduction: myths, mistakes, disbeliefs and confusion. , 2009, Reproductive biomedicine online.

[16]  A. Stavreus-Evers,et al.  Vitrification with DMSO protects embryo membrane integrity better than solutions without DMSO. , 2008, Reproductive biomedicine online.

[17]  A. Bakken Cryopreserving human peripheral blood progenitor cells. , 2006, Current stem cell research & therapy.

[18]  D. Mourelatos Chromosomes study as predictor of chemoresponse of tumours , 1996 .

[19]  A. Goldstone,et al.  Peripheral blood progenitor cells versus bone marrow. , 1994, Journal of hematotherapy.

[20]  C. Brayton Dimethyl sulfoxide (DMSO): a review. , 1986, The Cornell veterinarian.

[21]  Swanson Bn Medical use of dimethyl sulfoxide (DMSO). , 1985 .