Effect of liquid nitrogen vapor storage on the motility, viability, morphology, deoxyribonucleic acid integrity, and mitochondrial potential of frozen-thawed human spermatozoa.
OBJECTIVE
To evaluate the effectiveness of liquid nitrogen (LN(2)) vapor (indirect contact method) for the storage of human semen.


METHODS
Experimental study.


METHODS
University hospital-based fertility center.


METHODS
We evaluated 150 patients with normal sperm parameters.


METHODS
Human semen (N = 120) was mixed with Semen Freezing Medium, divided into two groups, frozen, and stored in LN(2) or LN(2) vapor. Frozen semen from each group (n = 40) was thawed after storage for 1 week, 1 month, or 3 months and then analyzed. In the second experiment, semen (n = 30) was divided into four groups, frozen, and stored in LN(2) or stored 7 cm, 12 cm, or 17 cm above the surface of LN(2) for 1 week.


METHODS
The motility and viability of sperm were evaluated by basic analysis, the morphology was analyzed with use of staining, the DNA integrity was assessed with use of terminal deoxyuridine triphosphate nick end-labeling assays, and active mitochondria were detected with use of rhodamine 123 staining.


RESULTS
The LN(2) and LN(2) vapor groups did not differ with regard to sperm motility, viability, morphology, DNA integrity, or active mitochondria when the cryostorage periods were compared. Furthermore, the quality of sperm stored within 17 cm of the surface of LN(2) for 1 week did not change.


CONCLUSIONS
The storage of human semen in LN(2) vapor, without direct contact with LN(2), may represent a useful alternative for the effective storage of human semen.
Lim
Jung Jin
JJ
Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul, South Korea.
Shin
Tai Eun
TE
Song
Seung-Hun
SH
Bak
Chong Won
CW
Yoon
Tae Ki
TK
Lee
Dong Ryul
DR
D000328:Adult; D048430:Cell Shape / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods; D053938:DNA Fragmentation / Q000187:drug effects*; D005615:Freezing / Q000009:adverse effects; D005740:Gases / Q000009:adverse effects / Q000494:pharmacology; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D009584:Nitrogen / Q000009:adverse effects / Q000737:chemistry / Q000494:pharmacology*; D044367:Phase Transition; D055101:Semen Analysis; D012662:Semen Preservation / Q000009:adverse effects / Q000379:methods; D013081:Sperm Motility / Q000187:drug effects*; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D016896:Treatment Outcome
D005740:Gases; D009584:Nitrogen