Magnetic-activated cell sorting for sperm preparation reduces spermatozoa with apoptotic markers and improves the acrosome reaction in couples with unexplained infertility.
BACKGROUND
Couples with unexplained infertility (UI) tend to have low fertilization rates with current IVF procedures. Here, we attempted to identify spermatozoa with apoptotic markers in couples with UI and unsuccessful intrauterine insemination (IUI) and we investigated the efficiency and benefit of magnetic-activated cell sorting (MACS) for sperm preparation in such patients.


METHODS
Sixty couples with UI and two IUI failures were recruited. The sperm were prepared by conventional density gradient centrifugation (DGC) and divided into two aliquots. One aliquot was used as a control and the other was further processed by MACS (D + M). Apoptotic markers were identified using fluorescence-labeled dye and flow cytometry, including externalization of phosphatidylserine (EPS), disrupted mitochondrial membrane potential (MMP) and DNA fragmentation. The fertilization potential of prepared spermatozoa was analyzed by basic semen analysis, computer-aided sperm analysis and the induced acrosome reaction test (IART).


RESULTS
After DGC, spermatozoa showed 18.6% EPS, 28.3% disrupted MMP and 13.5% DNA fragmentation. Numbers of spermatozoa with apoptotic markers were significantly reduced by D + M, versus DGC alone (P < 0.001). Although the motility of spermatozoa was slightly decreased after MACS, most sperm motion characteristics were not impaired. Interestingly, the IART significantly improved after D + M, versus DGC alone, especially for the couples with a normal hemizona assay (P < 0.001).


CONCLUSIONS
The spermatozoa prepared by D + M showed a reduced level of apoptotic markers. Improvement in the IART suggests a high fertilization potential of the processed spermatozoa. The identification of apoptotic markers and use of MACS may be helpful in directing the management plan for patients with UI and multiple IUI failures.
Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
Tsung-Hsien
TH
Lee
Chung-Hsien
CH
Liu
Yang-Tse
YT
Shih
Hui-Mei
HM
Tsao
Chun-Chia
CC
Huang
Hsiu-Hui
HH
Chen
Maw-Sheng
MS
Lee
D020101:Acrosome Reaction; D000328:Adult; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D002469:Cell Separation / Q000379:methods*; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007246:Infertility / Q000175:diagnosis / Q000628:therapy*; D007317:Insemination, Artificial, Homologous; D008280:Magnetics; D008297:Male; D011247:Pregnancy; D027724:Reproductive Techniques, Assisted; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism / Q000502:physiology
D017304:Annexin A5