• Clinical and Experimental Reproductive Medicine
• /
• v.33 no.1
• /
• pp.53-60
• /
• 2006

Objective: We tested the usefulness of swim-down technique using human follicular fluid (hFF) in sperm preparation. Methods: Semen samples were obtained from twelve male partners showing asthenozoospermia (sperm motility < 50%) at the time of routine andrologic evaluation in Seoul National University Bundang Hospital. After dividing into two aliquots, each samples were processed either by swim-down using 100% hFF or density gradient using SpermGrad. Sperm quality was assessed by computer-assisted semen analyzer (CASA). Results: Motility, Rapid motility, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement), and hyperactivated sperms were significantly increased, and LIN (mean linearity) was decreased significantly after sperm preparation in both groups. Motility was significantly higher after swim-down using 100% hFF when compared with density gradient using SpermGrad ($81.2{\pm}4.7$ vs. $67.6{\pm}2.3$, p=0.02) The other parameters assessed by CASA were not different between the two methods. Conclusion: Swim-down method with 100% hFF may be a useful method in preparation of sperm from asthenozoospermia.

• Clinical and Experimental Reproductive Medicine
• /
• v.35 no.3
• /
• pp.203-212
• /
• 2008

Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

• 대한생식의학회:학술대회논문집
• /
• 1995.12a
• /
• pp.68.1-68.1
• /
• 1995

• Corrugated packaging logistics
• /
• v.5 no.23
• /
• pp.107-115
• /
• 1998

• 대한생식의학회:학술대회논문집
• /
• 1999.11a
• /
• pp.49-50
• /
• 1999

• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.2
• /
• pp.225-232
• /
• 1997

폐색성 혹은 비폐색성 무정자증에서 부정소 정자채취법 등이 부적절하다고 여겨질때는 정소 조직을 일부 절제하여 그 조직으로부터 정자를 직접 채취하게 되는데 일반적으로 이렇게 정소로부터 추출한 정소정자는 운동성이 전혀 없거나 매우 약한 운동성을 보이는 경우가 많다. 본 연구의 목적은 이러한 정소정자를 Vero cell과 공배양을 시킴으로써 운동성을 획득시키거나 향상시키고 이를 수정시키는 시기까지 지속시킴으로써 정소정자추출술 (TESE)을 시행하는 환자나 의료진들에게 보다 편안하고 융통성있는 시간대를 부여하고, 아울러 정자직접주입술 (ICSI)을 보다 용이하게 하여 성공적인 수정률과 임신율을 얻음에 있다. 또한 ICSI를 시행한 후, 운동성이 향상된 잉여의 정소정자를 냉동보존함으로써 차후에 TESE을 다시 시행치않고도 시험관 아기 시술을 시도할 수 있는 부가적인 잇점도 있다고 할 수 있다. 대상환자군은 정관폐색증(n=11) 혹은 비정관폐색증(n=2)을 보이는 13명의 무정자증의 남성불임환자였으며 난자회수예정일 3일전에 TESE를 시행하여 정소정자를 얻은 후 이를 정자직접주입술이 시행되는 당일까지 Vero cell과 공배양을 실시하였다. Vero cell과의 공배양에 의하여 운동성이 있는 정소정자의 수는 공배양전과 비교하여 평균 3.3배가 증가하였으며, 특히 공배양전에 운동성이 있는 정소정자의 수가 50,000/ml이하의 미약한 운동성만을 보였던 경우 (n=5)에는 공배양 후에 운동성이 있는 정소정자 수의 평균증가율이 7.7배였다. 공배양전 정자운동성이 전혀 없었던 2례의 비정관폐색증환자중 3일간의 공배양을 통하여 1례에서 운동성을 획득한 정소정자를 얻을 수 있었으며 (14,300/ml), 정자직접주입술을 통하여 성공적인 수정 및 임신에 도달할 수 있었다. Vero cell과 공배양을 하고 ICSI했던 결과, 평균 수정률은 75.0% 이었으며 임신율은 61.5%였다.

• 대한생식의학회:학술대회논문집
• /
• 2006.11a
• /
• pp.123-124
• /
• 2006

• 대한생식의학회:학술대회논문집
• /
• 2007.11a
• /
• pp.102.2-103
• /
• 2007

• Development and Reproduction
• /
• v.2 no.1
• /
• pp.63-68
• /
• 1998

This study was done to find optimal cryopreservation medium and method to improve the post-thaw motility of human sperm. Thirty three semaen samples were included in the study. Of these, nineteen Samples showing normal semen profile were frozen using three cryprotectants (TYB, TYB+DTT and KS II)to compare post-thaw motility. Fourteen samples were frozen with vapor freezing and programmable freezing methods to compare post-thaw motility in correlation with freezing method. After 24 hrs of cryostorage , the vials were thawed and the post-thaw sperm motility was assessed by two kinds of computer-aededsperm analysis (CASA; SAIS and Hamilton Thorn). As a result, the post-thaw motility of the KS II group was higher than the TYB or TYB+DTT group in normal semen(34.8%, 28.3% and 23.0%, respectively). In the asthenospermia group, a significantly hither post-thaw motility was observed in the KS II (18.5%) compared to those of TYB or TYB+DTT group (13.6%, 10.0%, respectively). No difference was observed between vapor freezing group and computerized freezing group in normal semen (27.8%, 33.2%, respectively) and semen group showing asthenospermia (12.8%, 12.9%, respectively). In conclusion, our results indicate that KS II medium is reliable for cryopreservation of human sperm and vapor freezing and programmable freezing were equally effective in terms of the recovery of motile spermatozoa.

• Clinical and Experimental Reproductive Medicine
• /
• v.17 no.1
• /
• pp.71-80
• /
• 1990

New immunoassay systems for the detection of anti-sperm antibodies were developed. For this, sperm surface protein was purified by the immunoaffinity column prepared by the coupling of rabbit anti-human IgG antibodies to Sepharose-4B. Fraction eluted by tris-HCI buffer containing SDS showed a single band having molecular weight of about 60KD on electrophoresis. Enzyme HRP labelled goat anti-human IgG and chemiluminescence aminobutylethyl-isoluminol(ABEI) labelled rabbit anti-human IgG were used for ELISA and CIA, respectively. These two labelled conjugate bound well with human IgG. When serum dilution curves were made to titrate positive serums, two kinds of curves with steep and sluggish slopes were obtained Serum samples were categorized into 3 groups: positive, weak positive and negative based on slope of curve and O.D. values at 1:160 dilution of serum. When ELISA and CIA were compared to conventional method Kibrick test by the determinations of 62 male serums with different diagnosis, the results of ELISA and CIA agreed well, but both disagreed with that of Kibrick test. This study showed that purified sperm surface antigen can be used to develope solid-phase immunoassay systems such as ELISA and CIA which may eliminate the problems encounted the immobilization of living sperm in other tests.