Hi

Sperm cells storage and handling is important when doing in vitro or IVF, ICSI

temperature maintenance and perfect cryo medium or vitrification will helpful to maintain sperm cells live also it avoids the sperm DNA damage.

 Above said only suits for healthy (fertile) cells.  In case of infertile cells, cells damage will be their. 

Thank you very much Dear Ranganathan

Yes, this is true; however, if there any paper was working on this manner? In particular, in domestic animals. 

CryoLetters 24, 171-180 (2003)

Ó CryoLetters, c/o Royal VeterinaryCollege, London NW1 0TU, UK

EVALUATION OF DAMAGE IN

PACIFIC OYSTER (Crassostrea gigas) SPERMATOZOA

BEFORE AND AFTER CRYOPRESERVATION USING COMET ASSAY

Basic Clin Androl. 2016; 26: 11.

Published online 2016 Sep 27. doi:  10.1186/s12610-016-0038-3

PMCID: PMC5037879

Sperm DNA damage and assisted reproductive technologies: reasons to be cautious!

Joël R. Drevet

Dear Jyothi

Thank you very much 

I think the handling is not big problem in quality.

I am not sure if the handling in lab could affect sperm DNa fragmentation, but there are many information showing that  cryopreservation process per se could increase DNA fragmentation, related to ROS production.  

Dear respected Yong-Pil Cheon and Alexei Santiani 

Yes it is very true there are different factors could be the cause of DNA fragmentation such as genetic disorder, sperm Cryopreservation and other causes. 

For me I am writing a review related to Sperm DNA integrity and I want to cover all causes that might be the reason  for sperm DNA fragmentation.

Regards

Dear Falah Hasan,

• It is well known that all processes during handling of sperm cause DNA fragmentation. However, level of damage can be reduced some precautions during in vitro processes.
• We can say three factors which have detrimental effects on DNA during in vitro processes: 1) Light (sun light and especially florescent lambs because of UV), 2) temperature changes and 3) ROS especially H2O2 (H2O2, precursor of OH, is not considered as a ROS but it is well known that directly impairs DNA).
• Precautions are that you have to work in a dark side as possible as... If you need artificial light use halogen lamb or LED, and use colored tube or straws; you have to prevent heat changes and use cryoprotectant agents (egg yolk, milk based extenders, glyserol etc. or proper mixes of these agents) if it is difficult to prevent temperature changes; and you should complete process as possible as short period and use antioxidant agents against H2O2, i.e. catalase, GPx, superoxide dismutase etc. (these antioxidants prevent H2O2 production or scavenge H2O2).
• I hope these short information may be useful for you... 
• My best wishes,
• Umut  

I agree with Umut. Working with low oxygen incubation is also important to avoid oxydative stress, especially when spermatozoa are out of seminal plasma.

Also, there a some publications that shows that the technique of sperm washing is also a factor to consider as final fraction of spermatozoa present with different DNA damage intensity. Especially, type of selection technique (Swim up, DGC, simple washing, HA binding, MACS-Annexin5...) and presence/absence but also speed of centrifugation are factors to consider to avoid/reduce DNA fragmentation in the final sample.

Best regards,

Xavier