• 약학회지
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• 제50권6호
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• pp.409-414
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• 2006

Although mammalian sperms are cryopreserved for in vitro fertilization a process of cryopreservation decreases the fertility. Acrosome reaction requires depolarization-induced Ca$^{2+}$ influx and Ca$^{2+}$ releases from the Ca$^{2+}$ stores. To examine whether the cellular Ca$^{2+}$ mobilization is altered by a sperm cryopreservation we compared cytosolic Ca$^{2+}$ signals between fresh and cryopreserved pig sperms using confocal Ca$^{2+}$ imaging. The magnitudes of depolarization induced Ca$^{2+}$ increases were significantly smaller in cryopreserved sperms. Exposures to 10 mM caffeine or 5 ${\mu}$M thapsigargin elicited less Ca$^{2+}$ increases in the cryopreserved sperms compared to fresh sperms. In addition, progesterone-trig-gered Ca$^{2+}$ rises, that are thought to enhance acrosome reaction, were completely abolished in the cryopreserved sperms. These results suggest that storage and(/or) release of Ca$^{2+}$ from the intracellular Ca$^{2+}$ stores in pig sperms are significantly impaired by the process of cryopreservation.

• Fisheries and Aquatic Sciences
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• 제24권2호
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• pp.63-77
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• 2021

The aim of this research was to investigate different factors, including cryoprotective agents (CPAs), diluents, dilution ratios, equilibrium times, freezing rates, and thawing methods to optimize cryopreservation protocols for large yellow croaker (Larimichthys crocea). The parameters evaluated were sperm motility, sperm activity index (SAI), survival rate, and DNA damage. Different types of CPAs, such as dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), methanol, and glycerol, were tested for sperm preservation. The highest motility, SAI, and survival rate were observed when EG was used. Different diluents such as Stein's solution, Hank's balanced salt solution, marine fish Ringer's solution, artificial seminal plasma (ASP) of small yellow croaker, and Cortland solution were investigated. The highest post-thaw motility was observed upon using ASP as the diluent. Different concentrations of EG were then mixed with ASP to identify the optimal EG concentration. Experimental results showed that the motility (70.33 ± 1.20%), SAI (5), and survival rate (78.30 ± 0.42%) of post-thaw sperm were optimum when 10% EG and ASP were used as the CPA and diluent of cryopreservation, respectively. Post-thaw sperm motility was high at equilibration times below 150 s and at an optimum dilution ratio of 1:1 (sperm: CPA + diluent) and was not significantly different compared with fresh sperm motility. The freezing rate was found to be slow below -10℃/min. The thawing temperature of 45℃ was identified as ideal. The percentage of tail DNA in post-thaw sperm at 10% EG and ASP was also investigated and was found to have more significant DNA damage than that in fresh sperm but significantly lower damage than that in post-thaw sperm at EG concentrations of 5%, 15%, and 20% (p < 0.05). The cryopreservation protocols obtained in this study will be useful in large yellow croaker hatcheries.

• Animal Bioscience
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• 제36권10호
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• pp.1530-1535
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• 2023

Objective: Semen cryopreservation result in decreased sperm parameters and fertilization ability. Sericin exhibits antioxidant activity by reducing lipid peroxidation resulting from free radicals, which can potentially improve cryopreservation outcomes. The present study aimed to examine the efficacy of various sericin concentrations supplemented with a rooster semen-freezing extender on post-thaw semen quality and fertilizing ability of sperm after cryopreservation. Methods: Semen samples were collected from 40 roosters (5 reps), then were pooled, and divided into four groups by the levels of sericin supplementation (0%, 0.25%, 0.50%, and 0.75%) in a freezing extender. Semen suspensions were loaded in medium straw (0.5 mL) and cryopreserved with the traditional liquid nitrogen vapor method. Post-thawed semen was evaluated for sperm motility, sperm viability, and lipid peroxidation. Also, the fertility test was determined. Results: The results showed that supplementation of the freezing extender with 0.50% to 0.75% sericin resulted in greater total motility and progressive motility and lower malondialdehyde levels than the other groups after cryopreservation (p<0.05). However, the viability of 0.75% decreased compared with the value of 0.50% sericin supplementation (p<0.05). Moreover, the fertility and hatchability of total eggs were significantly higher in the 0.50% sericin group than in the other groups (p<0.05). Conclusion: In conclusion, 0.50% sericin is recommended as an alternative component of the freezing extender to improve cryopreserved rooster semen.

• 한국동물생명공학회지
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• 제35권3호
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• pp.268-272
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• 2020

Amniotic membrane stem cells are considered as a good alternative to embryonic stem cells, but their use in clinical studies is still not common. Here, exosomes from canine amniotic membrane mesenchymal stem cells (cAmMSC-exo) were used for dog sperm cryopreservation. Upon cryopreserved straws using cryoprotectant containing 0, 0.5, 1, or 2 ㎍/mL of cAmMSC-exo were thawed, motility and membrane integrity were analyzed. However, results showed no significant differences between the groups. We concluded that cAmMSC-exo with lower than 2 ㎍/mL have no effects on sperm cryopreservation, and further studies to get higher concentrations of cAmMSC-exo should be conducted for clinical application.

• Parasites, Hosts and Diseases
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• 제34권2호
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• pp.155-157
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• 1996

쥐와포자충(MCR주)을 장기간에 걸쳐 그 생명을 유지시키기 위하여 냉동 보존을 시도하였다. 그 오오시스트를 냉동보호제를 첨가하지 않고 증류수에 현탁시켜 $-20^{\circ}C$에서 24시간 예비냉각시킨 다음 $-70^{\circ}C$에 냉동보존하였다. 냉동 오오시스트를 $5^\circ}C$에서 해동시켜 마우스에 접종시킨 바 15개월간 냉동보존시켜토 사멸하지 않고 마우스에의 감염성이 유지되었다. 한편, 2.5% 중크롬산칼륨에 현탁시켜 6.5개월간 $5^{\circ}C$에 냉장한 오오시스트는 이미 사멸하여 감염성이 소실되었다.

• 한국작물학회지
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• 제49권4호
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• pp.354-357
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• 2004

A simplified technique which cryoprotects zygotic embryos by encapsulation-dehydration was developed for the germplasm conservation of herbaceous peony (Paeonia lactiflora Pall.). The highest survival rate $(85\%)$ was obtained from embryos treated by encapsulation-dehydration. The zygotic embryos were precultured on MS medium containing 0.3mg/L $GA_3$ for 1 day. The precultured embryos were encapsulated in $3\%$ (w/v) alginate beads and immersed for 1 h in MS medium containing 2 M glycerol and 0.5 M sucrose. The encapsulated embryos were dehydrated for 5h by air drying prior to direct immersion in liquid nitrogen. This encapsulation-dehydration method appears to be a promising technique for germplasm cryopreservation of a herbaceous peony.

• Journal of Chest Surgery
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• 제23권4호
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• pp.622-639
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• 1990

The use of aortic valve homograft has been developed since 1962 when Ross and Barratt - Boyes independently replaced a diseased aortic valve with an orthotopically inserted homograft valve. And also surgical treatment of complex congenital cardiac malformations utilizing homograft extracardiac conduit has been tried with better result than any other prosthetic material. The present study was undertaken to clarify the safety tissue viability, sterility, after following our protocol of procurement of heart, dissection of aortic and pulmonic homograft, sterilization, cryopreservation, thawing and dilution, and transplantation on experimental animal, sheep. Tissue viability of valve and great artery was assessed by tissue culture. Sterility was evaluated by bacterial and fungal culture. The method used was proven no deleterious effect on the integrity of the valve. Tissue culture of valve tissue before, and after cryopreservation process resulted that active fibroblast growth was observed from homograft sterilized with antibiotics. And culture of the transplanted homograft from sacrificed animal showed active fibroblast growth. Pathologic examination of implanted valve tissue from sacrificed sheep showed mild calcification and minor change, but there were moderate and severe calcification of wall of great arteries.

• 한국어류학회지
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• 제19권2호
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• pp.173-177
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• 2007

참돔 정자의 동결보존에 미치는 희석제와 동해방지제의 효과를 운동성과 생존율로 비교하였다. 300 mM glucose를 희석제로 사용하였을 때 보존 효과가 좋았다. 동해방지제로는 10% dimethyl sulfoxide (DMSO)를 동결보존에 사용하였을 때 해동 후의 운동성과 생존율이 가장 높았다.

• Fisheries and Aquatic Sciences
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• 제13권4호
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• pp.350-353
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• 2010

This study demonstrated that cryopreserved semen from dead fish can be used for seedling production. Yellow croakers, Larimichthys polyactis, were killed and stored at temperatures of $20^{\circ}C$ or $0^{\circ}C$ for 6 hours. At 2 hour intervals, semen from these fish was collected using abdominal pressure and evaluated for spermatozoa motility and semen cryopreservation. Semen collected after 6 hours from dead fish stored at $0^{\circ}C$ could be cryopreserved and attained fertilization and hatching rates of $15.0{\pm}1.2%$ and $14.8{\pm}1.6%$, respectively. This study suggests that germ cells such as the semen of dead fish can be cryopreserved and utilized in the restoration of a species.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.325-330
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• 1994

The study has been carried out in order to evaluate the effects of embryonic stage, and cryopreservation method on the rates of viability and development of the cryopreserved mouse early embryos. The results were as following:In the treatment steps of cryoprotectant, for the fertilized oocyte with pronucleus(PN), 2-step was better than the others. And for the other embryos, 4-step was better than 2- or 3-step. In respect to the embryonic stage, as the embryos developed from fertilized oocytes to 8-cell embryos, the rates of viability and development were increased higher. Therefore, 8-cell embryo was better stage than the others. In respect to the kind of cryoprotectants, PROH was better than DMSO for the fertilized oocyte, as a cryoprotectant. DMSO, for the 2-cell embryos and PROH and DMSO for the 4- and 8-cell embryos were suitable for cryopreservation.