PMID- 11451362
OWN - NLM
STAT- MEDLINE
DCOM- 20011207
LR  - 20041117
IS  - 0196-3635 (Print)
IS  - 0196-3635 (Linking)
VI  - 22
IP  - 4
DP  - 2001 Jul-Aug
TI  - Cryopreservation-Thawing of fractionated human spermatozoa is associated with 
      membrane phosphatidylserine externalization and not DNA fragmentation.
PG  - 646-51
AB  - The objective of these studies was to evaluate the effect of 
      cryopreservation-thawing of human spermatozoa on DNA fragmentation and membrane 
      integrity. This was a prospective, controlled cohort study, performed at a 
      university-based infertility center. Ejaculates were examined from 5 donors and 
      16 men undergoing infertility evaluation. Purified sperm populations were 
      prepared by gradient centrifugation, cryopreserved using a manual method and 
      TEST-yolk buffer and glycerol (TYB-G), followed by quick-thaw. Annexin V binding 
      was used for assessing membrane translocation of phosphatidylserine, and terminal 
      deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) was utilized 
      for the evaluation of DNA fragmentation. The results were as follows: the 
      percentage of live cells with intact membranes (annexin V-, live) was 
      significantly reduced after cryopreservation-thawing. On the other hand, the 
      percentages of live cells with phosphatidylserine translocation (annexin V-, 
      live) and of necrotic (dead) cells increased significantly after thawing. TUNEL 
      revealed percentages of cells with DNA fragmentation in the prefreeze and 
      postthaw samples that were not significantly different. In a further attempt to 
      examine differences in response to various cryoprotection protocols, experiments 
      were carried out using no cryoprotection, glycerol alone, or TYB-G. Samples 
      frozen with TYB-G demonstrated significantly higher percentages of live cells 
      without phosphatidylserine translocation than the other conditions. We concluded 
      that cryopreservation-thawing of human sperm from patients and donors was 
      associated with membrane change, as revealed by membrane translocation of 
      phosphatidylserine, while having no major impact on DNA fragmentation.
FAU - Duru, N K
AU  - Duru NK
AD  - The Jones Institute for Reproductive Medicine, Department of Obstetrics and 
      Gynecology, Eastern Virginia Medical School, Norfolk 23507, USA.
FAU - Morshedi, M S
AU  - Morshedi MS
FAU - Schuffner, A
AU  - Schuffner A
FAU - Oehninger, S
AU  - Oehninger S
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Androl
JT  - Journal of andrology
JID - 8106453
RN  - 0 (Annexin A5)
RN  - 0 (Phosphatidylserines)
SB  - IM
MH  - Annexin A5/metabolism/pharmacology
MH  - Cell Fractionation
MH  - Cell Membrane/chemistry/metabolism
MH  - Cryopreservation
MH  - *DNA Fragmentation
MH  - Humans
MH  - In Situ Nick-End Labeling
MH  - Male
MH  - Phosphatidylserines/analysis/*metabolism
MH  - *Semen Preservation
MH  - Sperm Motility
EDAT- 2001/07/14 10:00
MHDA- 2002/01/05 10:01
CRDT- 2001/07/14 10:00
PHST- 2001/07/14 10:00 [pubmed]
PHST- 2002/01/05 10:01 [medline]
PHST- 2001/07/14 10:00 [entrez]
PST - ppublish
SO  - J Androl. 2001 Jul-Aug;22(4):646-51.