• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.52-55
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• 2017

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

• Clinical and Experimental Reproductive Medicine
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• 제46권1호
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• pp.36-40
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• 2019

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients' semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.

• 한국발생생물학회지:발생과생식
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• 제3권1호
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• pp.39-44
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• 1999

성선자극호르몬으로 자극된 난소로부터 회수된 사람의 성숙난자의 전핵형성과 초기발생에 정자의 운동성과 성숙도가 미치는 영향을 조사하였다. 세포질내정자주입 (ICSI)은 HEPES-buffer mTCM-199 배양액에서 실시하였다. 체내에서 성숙된 난구세포부착난자를 ICS에 의하여 사정된 운동성 정자 또는 비운동성 정자로 수정하였을 때 운동성 정자로 수정된 난자가 비운동성 정자로 수정된 난자보다 전핵형성율이 높았다 (79.8% vs 51.7% ; p<0.002)). 그러나 체내에서 성숙된 난구세포부착난자를 ICS에 의해 정소 내 운동성 정자 또는 비운동성 정자로 수정하였을 때 운동성 정자와 비운동성 정자 사이의 전핵형성율은 차이가 없었다. 10.0 mM lactate, 0.5 mM pyruvate, 0.2 mM taurine, 1.0 mM glutamine, 2.22 mM MEM amino acids, vitamin 그리고 10% 사람 난포액이 포함된 수정 Tyrode 배지에서 전핵이 형성된 수정란의 초기 발생은 정자의 채취원과 운동성에 관계없이 9∼16세포기로 발생하였다.

• Tuberculosis and Respiratory Diseases
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• 제53권4호
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• pp.457-462
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• 2002

저자들은 우연히 발견된 Kartagener 증후군의 삼주징을 가진 여자환자에서 기관지점막의 섬모에 대한 전자현미경적 미세구조 분석을 시행하여 한 명의 환자에서 내, 외측 dynein arm의 결함, radial spoke의 결함, microtubule의 전위, 거대섬모 등 네 가지 이상의 다양한 미세구조결함이 혼재하는 비교적 드문 환자를 경험하였기에 보고하는 바이다.

• 한국수산과학회지
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• 제30권5호
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• pp.809-815
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• 1997

문치가자미의 성숙한 수컷 15마리를 재료로 하여 삼투질농도와 $Ca^{2+}$이 정자의 운동성에 미치는 영향을 파악하였다. 문치가자미 정자는 전해질 및 비전해질 용액에서 정장의 삼투질농도인 $351{\pm}23.4mOsm/kg$ 이하의 저장액에서 운동이 억제되었으나, 삼투질농도 $466\~l,185mOsm/kg$에서는 활발한 운동을 보였으며, 1,698 mOsm/kg 이상에서 다시 운동이 중지되었다. 인공해수를 희석액으로 하여 pH를 달리하였을 때, 정자는 $pH\;5\~9$에서 운동지수 $3.75\~4.00$로 운동성이 활발하였으나, pH 4에서는 운동지수 2.50로 미약하였다. $Ca^{2+}$의 특이 chelator인 EGTA가 포함된 $Ca^{2+}$ 결여 인공해수 (tris-HCl, pH 7.4)에 정액을 희석하였을 때, EGTA의 농도 증가에 따라 정자 운동성이 낮아졌으며, 3mM 이상의 농도에서는 완전히 정지되었다. 운동이 억제된 정자에 $Ca^{2+}$을 서서히 첨가한 결과, $Ca^{2+}$ 농도 0.25mM 이상부터 운동이 재개되었다. 또한 인공해수속의 정자는 calmodulin 억제제인 TFP의 농도의 증가에 따라 운동성이 감소하여 0.5mM에서 완전히 정지하였다.

• 한국어류학회지
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• 제19권2호
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• pp.168-172
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• 2007

졸복의 정액의 물리화학적 성상과 정자운동성에 관한 기초자료를 얻었다. 졸복 정액의 평균농도, pH 및 삼투질농도는 각각 $12.1{\pm}3.2{\times}10^9/mL$, $8.2{\pm}0.3$, $385.5{\pm}12.5mOsm/kg$ 였다. 정자는 정장의 삼투질농도($385.5{\pm}12.5mOsm/kg$)에 비해서 저장 또는 등장의 삼투질농도에서는 운동성이 억제된 반면, 장장에 비해 고장삼투질농도에서 정자의 운동이 개시되어 삼투질농도가 증가함에 따라 활발한 운동성을 보였다.

• 한국임상수의학회지
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• 제24권4호
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• pp.577-583
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• 2007

It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

• Clinical and Experimental Reproductive Medicine
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• 제27권3호
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• pp.267-273
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• 2000

Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.