DNA integrity is maintained after freeze-drying of human spermatozoa

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Presentation transcript:

DNA integrity is maintained after freeze-drying of human spermatozoa Luca Gianaroli, M.D., Maria Cristina Magli, M.Sc., Ilaria Stanghellini, Ph.D., Andor Crippa, Ph.D., Anna Maria Crivello, B.Sc., Edoardo Stefano Pescatori, M.D., Anna Pia Ferraretti, M.D. Fertility and Sterility Volume 97, Issue 5, Pages 1067-1073.e1 (May 2012) DOI: 10.1016/j.fertnstert.2012.02.014 Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Sperm membrane integrity as assessed by the vital dye eosin in rehydrated spermatozoa. The dye penetrated in all cells, indicating membrane damage and lack of viability. Sperm head morphology seems to be well preserved, but tail curling is present in four cells. A sperm cell with two heads and two tails can be seen in the center of the figure. Fertility and Sterility 2012 97, 1067-1073.e1DOI: (10.1016/j.fertnstert.2012.02.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Evaluation of sperm indices before (triangles) and after (squares) lyophilization. Normozoospermic samples are those indicated with the numbers 1–19, and those with moderate teratozoospermia correspond to numbers 20–30. (A) After lyophilization, head morphology was preserved. (B) The proportion of spermatozoa with tail anomalies was statistically significantly increased. (C) The result was a statistically significant decrease in normal forms. Fertility and Sterility 2012 97, 1067-1073.e1DOI: (10.1016/j.fertnstert.2012.02.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Comparison of sperm head birefringence properties before (triangles) and after (squares) lyophilization. No statistically significant variations detected in (A) the proportions of spermatozoa with total birefringence, (B) birefringence localized in the postacrosomal region, and (C) interrupted birefringence due to the presence of vacuolelike structures. (D) Increased frequency of nonbirefringent sperm cells after rehydration. Fertility and Sterility 2012 97, 1067-1073.e1DOI: (10.1016/j.fertnstert.2012.02.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions

Supplemental Figure 1 The characteristics of birefringence were maintained after the freeze-drying procedure. (A) Completely birefringent head. (B) Partial birefringence localized in the lower part of the head consequent to the occurrence of the acrosome reaction. (C) Partial birefringence due to the presence of a vacuole-like structure. Fertility and Sterility 2012 97, 1067-1073.e1DOI: (10.1016/j.fertnstert.2012.02.014) Copyright © 2012 American Society for Reproductive Medicine Terms and Conditions