• Journal of Marine Life Science
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• v.8 no.2
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• pp.115-120
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• 2023

This study aims to find out a suitable extender and cryoprotective agent (CPA) for cryopreservation and its optimum concentration in order to conduct planned artificial seed production of Korean bullhead Pseudobagrus fulvidraco and to preserve superior sperm. Experiments were designed to investigate the effects of the different combinations of three extenders (I: 300 mM glycose, II: Kurokura extender, III: Li extender), four cryoprotectants (dimethyl sulfoxide, ethylene glycol, methanol and glycerol) and four concentrations (5, 10, 15, 20%) on the cryopreservation of Korean bullhead sperm. Postthawed sperm survival rate and sperm activity index (SAI) were detected to evaluate the effects of sperm cryopreservation. The optimal combination of extender and CPA for cryopreservation of Korean bullhead sperm was extender III + 10 and 15% methanol, resulting in a survival rate and SAI of 66.9 ± 8.7, 67.3 ± 13.1% and 2.6 ± 0.4, 2.6 ± 0.5 respectively, which was higher than had been achieved with other extenders and CPAs.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.81-81
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• 2003

돼지 정액의 동결보존은 돼지 정자가 내동성이 약한 특성을 가지고 있어 아직 만족할만한 결과를 얻지 못하여 액상정액을 이용하여 인공수정을 실시하고 있는 실정이다. 돼지 정액의 동결보존을 효과적으로 이룩한다면 돼지 인공수정의 효율성을 증대시킬 수 있고, 우수한 종돈의 보호 및 국제적 돼지 품종의 교류를 증진시킬 수 있을 것이다. 본 실험은 돼지 정액 동결시 동결온도와 시간, 항산화제로 알려진 Taurine의 농도별 첨가, 동해보호제 및 융해조건이 돼지 정액의 동결 융해후 생존성에 미치는 영향을 검토하고자 수행되었다.

• Journal of Sericultural and Entomological Science
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• v.37 no.2
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• pp.186-187
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• 1995

동결보존에서 가장 재생율이 높은 품종은 조직배양에서 발근력이 뛰어난 것으로 알려져 있는 신광뽕으로 최고 92.0%의 재생율을 보였으나, 개량, 용천 및 검설뽕에서는 재생을 보이지 않았다. 또, 건조과정을 거치지 않은 것이나, 건조과정과 예비동결 과정을 거치지 않고, 동결방어제만을 사용한 것의 재생은 전혀 확인할 수 없었다. 금후, 동결보존한 동아의 재생에서 돌연변이 유무는 RAPD로 확인될 수 있을 것이다.

• Korean Journal of Ichthyology
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• v.31 no.4
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• pp.195-200
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• 2019

An experiment was performed to obtain cryopreservation techniques of Pacific cod Gadus macrocephalus sperm. Milt were cryopreservation using five cryoprotectant demethyl sulfoxide (DMSO), ethylene glycol (EG), glycerol, methanol and propylene glycol (PG) with marine fish ringer's solution (MFRS) as diluent. Milt were cryopreserved each experimental methods like cryoprotectants (10% and 20%), equilibration time (3, 5 and 10 min) and freezing protocols (liquid nitrogen vapor above 3, 8 and 12 cm). Post-thaw sperm survival rate revealed the highest in 10% PG with optimum methods of equilibration time (3 min) and freezing protocol (liquid nitrogen vapor above 8 cm) about 21.3±1.8%. Hatching rate of fertilization eggs using fresh and cryopreserved sperm were no significantly different.

• Korean Journal of Ichthyology
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• v.19 no.2
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• pp.173-177
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• 2007

A series of experiments were conducted to compare the effects of various diluents and cryoprotectants on the motility and survival rate in cryopreservation of the red seabream, Pagrus major sperm. Sperm was efficiently cryopreserved using 300 mM glucose as a diluent. Two cryoprotectant, dimethyl sulfoxide (DMSO) and glycerol, were added to 300 mM glucose to formulate the extenders at concentrations between 5% and 30% by volume for freezing. The highest post-thawed sperm motility and survival rate were obtained with 10% DMSO.

• The Korean Journal of Malacology
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• v.26 no.4
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• pp.255-260
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• 2010

This study was conducted to investigate protocol standardization for spermatozoa cryopreservation of the Scapharca broughtonii (Schrenck). Among the freezing rates, freezing at a height of 2 cm above liquid nitrogen surface for 5 minute gave higher activity and survival rate. Among the various diluents, Ringer's solution was the best for S. broughtonii sperm cryopreservation. The suitability of cryoprotectants dimethylsulfoxide (DMSO), dimethylacetamide (DMA), glycerol and methanol were tested against three freezing rates. DMSO gave significantly higher activity and survival rates than others.

• KSBB Journal
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• v.21 no.2
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• pp.110-117
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• 2006

During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing serum with 10% DMSO. But, in case of bioprocess under the serum-free conditions, including cultivation of cell lines and producing of pharmaceuticals, the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the serum-free cryopreservation on the CHO cells. To improve the survival rates of the cryopreserved CHO cells in serum-free condition, first, the effects of permeable and non-permeable additives for substitute serum on cell viability were investigated. The combination of 10% DMSO and 0.03 M raffinose in MEM-${\alpha}$ without serum indicated 76% of cell viability. However, it did not reach the survival rates(more than 95%) of the conventional cryopreservation. In the second, to evaluate the cryopreservative ability of the serum-free medium(SFM) we compared viability of the CHO cells cryopreserved in the SFMs(Sigma C5467, C4726, and C1707, JBI SF486 and PF486), the cryoprotectant(Genenmed CAN-1000) and the MEM-${\alpha}$ with serum. All solution contained 10% DMSO. As a result of the comparison, cryopreserved cells in the SFMs showed over 95% of viability and appeared predominant viability better than cryoprotectant CAN-1000. Finally, we assessed the stability of the CHO cells in the long-term cryopreservation(LTC) using SFM. Every three months, the cryopreserved CHO cells were thawed to estimate the cell viability and the recovery rates. Then, real-time RT-PCR analyzed the inserted CHO DHFR gene. All results for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion, it could be seen that the LTC in the SFM can substitute for serum using methods in the bioprocess proceeded by CHO cells for more than 18 months.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.99-106
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• 1999

핵 이식에 공여되는 수핵난자의 효과적인 동결보존을 위하여 한우 성숙난자를 1.0 M dimethylsulfoxide(DMSO) 또는 1.0 M glycerol이 함유된 동결보호제를 이용하여 처리하거나, 동결보호제 처리 후 완만동결법을 이용한 동결융핼르 시행하여 상기 실험처리로 야기되는 투명대 경화현상을 관찰하였다. 도축장 유래의 난소에서 미성숙난자를 채취한 후 10% 소 태아혈청을 함유한 TCM-199을 이용하여 22∼24 시간 동안 체외성숙배양을 이해하였다. 배양후 작출된 성숙난자를 각각의 동결보호제로 처리, 혹은 처리 후 동격융해한 후 protease를 이용하여 투명 대의 경화현상 발생의 빈도를 조사하였다. 또한 동격란을 동결정액을 이용한 체외수정에 공여한 후 정자 침입농도능을 조사하였다. 동결보호제로 처리한 난자에 있어서 보호제의 종류와 관계없이 투명대 경화현상이 유의적 (P<0.05) 으로 증가하였으나 이후의 동결융해 처리에 의한 추가적인 경화현상의 발생은 증가하지는 않았다. 또한 투명대 경화현상의 발생양상을 동결보호제 처리 후 10분 간격으로 측정한 결과 DMSO의 경우 처리후 10분, glycerol의 경우 처리 후 20분 후부터 유의적으로 차를 발견할 수 없었으며, 수정율 및 난자 1개당 침입한 정자의 수는 동결란에서 유의적으로 증가하지 않았다. 본 연구의 결과 동결난자의 투명대 경화현상은 동결보호제 처리과정에서 이미 일어나지만, 이러한 투명대 경화현상이 난자의 동결보존 후 수정능에는 현자한 영향을 미치지 않는다는 사실이 규명되었다.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.34-35
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• 1998

• 대한생식의학회:학술대회논문집
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• 2005.06a
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• pp.107.2-108
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• 2005