PMID- 33830681
OWN - NLM
STAT- MEDLINE
DCOM- 20220112
LR  - 20220112
IS  - 2047-2927 (Electronic)
IS  - 2047-2919 (Linking)
VI  - 9
IP  - 4
DP  - 2021 Jul
TI  - Simultaneous detection of sperm membrane integrity and DNA fragmentation by flow 
      cytometry: A novel and rapid tool for sperm analysis.
PG  - 1254-1263
LID - 10.1111/andr.13017 [doi]
AB  - BACKGROUND: Sperm DNA integrity has become one of the most discussed and 
      promising biomarkers for the assessment of male fertility. However, an 
      easy-to-apply method capable of estimating DNA fragmentation in the live fraction 
      of spermatozoa has remained elusive, preventing this parameter from being fully 
      applied in clinical settings. OBJECTIVES: To validate a novel co-staining for the 
      analysis of DNA fragmentation in membrane-intact spermatozoa. MATERIALS AND 
      METHODS: Normozoospermic semen samples were used to validate the co-staining 
      consisting of acridine orange (AO) and LIVE/DEAD Fixable Blue Dead Cell Stain 
      (LD), against established methods for the evaluation of cell viability, propidium 
      iodide stain (PI), and DNA fragmentation, the sperm chromatin structure assay 
      (SCSA), to rule out cross-interference. Furthermore, the accuracy of the method 
      was tested by the evaluation of samples prepared with different amounts of 
      membrane and DNA damage (20, 40, 60, 80, and 100%). RESULTS: No significant 
      differences were observed between the co-staining and the established staining 
      procedures (membrane integrity, p = 0.755; DNA fragmentation p = 0.976). 
      Moreover, high R square values were obtained from the analysis of samples of 
      known membrane (R(2)  = 0.9959) and DNA damage (R(2)  = 0.9843). The simultaneous 
      assaying of sperm membrane integrity and nuclear DNA fragmentation allowed the 
      analysis of four sperm categories and thereby to assess the proportion of 
      membrane-intact spermatozoa with compromised DNA integrity. DISCUSSION AND 
      CONCLUSION: This new protocol has the potential to provide clinically relevant 
      information about the DNA fragmentation in membrane-intact spermatozoa. Thus, it 
      has the potential of improving the diagnostic of male infertility and enabling a 
      better understanding of sperm dysfunction.
CI  - (c) 2021 American Society of Andrology and European Academy of Andrology.
FAU - Da Costa, Raul
AU  - Da Costa R
AD  - Centre for Reproductive Medicine and Andrology, University of Munster, Munster, 
      Germany.
FAU - Redmann, Klaus
AU  - Redmann K
AD  - Centre for Reproductive Medicine and Andrology, University of Munster, Munster, 
      Germany.
FAU - Schlatt, Stefan
AU  - Schlatt S
AUID- ORCID: 0000-0002-7571-5313
AD  - Centre for Reproductive Medicine and Andrology, University of Munster, Munster, 
      Germany.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210504
PL  - England
TA  - Andrology
JT  - Andrology
JID - 101585129
SB  - IM
MH  - Adult
MH  - *DNA Fragmentation
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - Infertility, Male/diagnosis
MH  - Male
MH  - Semen Analysis/*methods
MH  - *Spermatozoa
MH  - Staining and Labeling/*methods
OTO - NOTNLM
OT  - DNA damage
OT  - acridine orange
OT  - flow cytometry
OT  - membrane integrity
OT  - sperm DNA fragmentation test
EDAT- 2021/04/09 06:00
MHDA- 2022/01/13 06:00
CRDT- 2021/04/08 13:01
PHST- 2021/01/19 00:00 [revised]
PHST- 2020/11/19 00:00 [received]
PHST- 2021/03/05 00:00 [accepted]
PHST- 2021/04/09 06:00 [pubmed]
PHST- 2022/01/13 06:00 [medline]
PHST- 2021/04/08 13:01 [entrez]
AID - 10.1111/andr.13017 [doi]
PST - ppublish
SO  - Andrology. 2021 Jul;9(4):1254-1263. doi: 10.1111/andr.13017. Epub 2021 May 4.