• 한국수산과학회지
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• 제55권5호
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• pp.670-680
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• 2022

Owing to the lack of a cold-chain distribution system, most seafood is generally distributed under room temperature conditions. However the degradation of freshness during the distribution process can lead to disputes between sellers and consumers. The most widely used method for low-temperature distribution for seafood includes packaging it with styrofoam boxes and cold packs. In this study, vacuum-packed frozen fillets of four fish species of [white meat (Paralichthys olivaceus and Sebastes schlegelii) and red meat (Scomber japonicus and Scomberomorus niphonius)] were placed in styrofoam boxes with cold packs. Thereafter, changes in chemical (including pH, volatile basic nitrogen, and trimethylamine), physical (odor intensity, hardness, and chewiness), and microbial (viable cell count) characteristics of the fillets were measured during storage at 25℃. To identify the suitable method of determining freshness during the room-temperature distribution, several factors were considered, which included significant difference verification, correlation coefficients, and economic efficiency (experimental cost and time). Volatile basic nitrogen, pH, odor intensity, and viable cell count are the most rapid and accurate freshness indicators for determining freshness of frozen fish fillets during.

• 한국가축번식학회지
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• 제17권3호
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• pp.201-207
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• 1993

We determined whether the ultrarapid freezing method is applicable to micromanipulated mouse embryos. One-cell mouse embryos were microinjected with MThGH gene. Nuclei from one-cell embryos of F1(C57BL$\times$CBA) mice were transplanted into enucleated one-cell embryos of ICR mice. The injected and nucleated embryos that developed to 2-cell stage were cryopreserved by ultrarapidfreezing. The embryos equilibrated in freezing medium(3 M DMSO＋0.25 M sucrose＋2% FBS in PBS) were directly immersed into liquid nitrogen and then thawed in 37$^{\circ}C$ water. Development rates of the microinjected and nuclear-transplanted embryos to blastocyst stage after ultrarapidly freezing and thawing were 31% and 55%, respectively. The frozen-thawed embryos were transferred to pseudopregnant recipient, which then gave birth to 17 offsprings. Twelve(14% of the transferred embryos) and five(20%) offsprings were derived from microinjected and nuclear-transplanted embryos, respectively. The results indicate that the DNA injected and nuclear-transplanted mouse embryos are cryopreservable at 2-cell stage by ultrarapid freezing method.

• Clinical and Experimental Reproductive Medicine
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• 제50권4호
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• pp.213-222
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• 2023

Cryopreservation is an option for the preservation of pre- or post-pubertal female or male fertility. This technique not only is beneficial for human clinical applications, but also plays a crucial role in the breeding of livestock and endangered species. Unfortunately, frozen germ cells, including oocytes, sperm, embryos, and spermatogonial stem cells, are subject to cryoinjury. As a result, various cryoprotective agents and freezing techniques have been developed to mitigate this damage. Despite extensive research aimed at reducing apoptotic cell death during freezing, a low survival rate and impaired cell function are still observed after freeze-thawing. In recent decades, several cell death pathways other than apoptosis have been identified. However, the relationship between these pathways and cryoinjury is not yet fully understood, although necroptosis and autophagy appear to be linked to cryoinjury. Therefore, gaining a deeper understanding of the molecular mechanisms of cryoinjury could aid in the development of new strategies to enhance the effectiveness of the freezing of reproductive tissues. In this review, we focus on the pathways through which cryoinjury leads to cell death and propose novel approaches to enhance freezing efficacy based on signaling molecules.

• 한국가축번식학회지
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• 제8권2호
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• pp.116-121
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• 1984

These experiments were carried out to examine the effect of rapid thawing (500$^{\circ}C$/min) on the survival of 8-cell mouse embryos cooled slowly (0.5-1$^{\circ}C$/min) to precooling temperatures between -10 and 070$^{\circ}C$ before direct transfer ofembryos to -196$^{\circ}C$, and to compare the survival of embroys thawed slowly (20$^{\circ}C$/min) and rapidly (500$^{\circ}C$/min) after in vitro culture. In addition, the sensitivity of 8-cell mouse embroys to the rate of addition and removal of cryoprotectant at room temperauture, and the effect of developing stages on the survival of embryos frozen-thawed slowly were investigated. The results obtained were summarized as follows: 1. Embryos were survived in rapid thawing (500$^{\circ}C$/min) only when slow cooling was terminated at relatively high subzero teperaure (-20 to -60$^{\circ}C$). The highest survival rate(77.0%) in in vitro culture of embryos thawed rapidly was obtaeined after transfer to -196$^{\circ}C$ from -40$^{\circ}C$. 2. For the survival of embryos in slow thawing (20$^{\circ}C$/min.), slow cooling to lower subzero temperature (-50$^{\circ}C$ and below) was required before transfer of embryos to -196$^{\circ}C$. These results indicate that embryos transferred to -196$^{\circ}C$ from high subzero temprature contain much interacellular ice to damage them during slow warming but to permit survival of embryos after rapid warming. 3. The Survival rate (72.7%) of 8-cell mouse embryos after rapid addition and removal of cryoprotectant, DMSO at room temperature was similar to that (83.9%) after slow addition and removal of cryoprotectant at same temperature. 4. The survival rates of 1-, 2-, 4- and 8-cell embryos frozen-thawed slowly were 26.7, 76.4, 70.0 and 83.9%, respectively.

• 한국수정란이식학회지
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• 제28권1호
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• pp.41-48
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• 2013

This study examined the motility of either the unattached(upper) or attached(lower) Hanwoo sperm to bovine oviduct epithelial cell(BOEC) monolayers to determine whether there are any changes in their motility during co-culture. The cleavage and blastocyst development rate were compared among different preincubation methods in-vitro, after oocytes were fertilized in-vitro with Hanwoo sperm on BOEC monolayers. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 5 hours and 6 hours (p<0.05) of incubation, in sperm treatment medium without heparin and caffeine. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 3 hours (p<0.05) and 6 hours (p<0.01), in sperm treatment medium containing heparin and caffeine. The motility of the attached( lower) sperm was significantly higher than the unattached(upper) sperm during co-culture with BOEC at all times(p<0.01 or p<0.05), except for 6 hours. After Hanwoo oocytes were fertilized in-vitro with the sperm that had been co-cultured with BOEC in sperm treatment medium containing heparin and caffeine, we determined the cleavage and blastocyst development rate, according to the preincubation methods. Both the cleavage and blastocyst development rate from 2 hour preincubation group were the highest, but significant difference was not recognized. These results show that BOEC plays an important role on sperm hyperactivation related to capacitation regardless of heparin and caffeine in sperm treatment medium. However, oviduct epithelial cell had no significant effect on the development of embryos after in-vitro fertilization in the presence of added heparin and caffeine in sperm treatment medium.

• Journal of Microbiology
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• 제41권3호
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• pp.262-265
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• 2003

Although enumerating bacterial cells is a fundamental step in understanding microbial ecosystems in marine environments, substantial decrease in bacterial counts with increasing sample storage time hampers the accurate estimation of bacterial biomass. We compared the variations in bacterial cell numbers caused by freezing and thawing of sample bottles or slides. Bacterial counts of seawater samples frozen only once in a sampling bottle yielded approximately 95％ of the original numbers after 90 days, whereas 80％ of the original count was obtained for samples prepared on slides. Only 67％ and 58％ of the original counts were recovered in samples repeatedly frozen and thawed in bottles or on slides, respectively. The results indicated that freezing a seawater sample in a bottle increased the consistency of the epifluorescence microscopic enumeration of bacterial cells.

• 대한수의학회지
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• 제31권2호
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• pp.229-234
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• 1991

Thc chimeric morulae were produced following aggregation of the half embryos which were microsurgically bisected at 8-cell and early morula stage. Different phenotypic embryos were obtained by mating ICR female mice with ICR or CBA male mice. The early morula stage was thc desirable stage for the aggregation of mouse embryos after bisection. The post-thawed survival rates of bisected-aggregated embryos that developed into normal blastocyst after conventional freezing in DMSO and ethylene glycol were 30.5 and 32.896, respectively. One offspring was produced by transferring the 67 frozen-thawed bisected-aggregated embryos.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.6-10
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• 2001

No. Sperm can be sexed with 90% accuracy by flow cytometry/cell sorting. No. The current speed of sexing is about 5,000 live sperm of each sex per second, remarkably fast considering that each sperm is individually sexed. No. Although fast, sperm sexing is not fast enough to use standard numbers of sperm per AI dose. No. With well managed heifers, pregnancy rates with low doses of sexed, frozen sperm are 70-80% of those with unsexed sperm with normal sperm numbers. Pregnancy rates are lower in lactating dairy cows. No. Calves from sexed sperm appear to be normal. No. Sexed, frozen semen from a few bulls currently is available commercially in the United Kingdom, and likely will be available in several other countries in 2002, probably at a premium of US ＄30-50 per straw. (omitted)

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1996년도 춘계학술대회
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• pp.119-130
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• 1996

We review the factors which regulate the depth and composition of the human airway surface liquid (ASL). These include secretion from airway submucosal glands, ion and fluid transport across the surface epithelium, goblet cell discharge, surface tension and transepithelial gradients in osmotic and hydrostatic pressure. We describe recent experiments in which we have used low temperature scanning electron microscopy of rapidly frozen specimens to detect changes in depth of ASL in response to submucosal gland stimulation. We also present preliminary data in which X-ray microanalysis of frozen specimens has been used to determine the elemental composition of ASL.

• 한국미생물·생명공학회지
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• 제20권4호
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• pp.470-476
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• 1992

Nocardia mediterranei NCK-4를 동결건조시 생존도에 미치는 영향을 검토하기 위해 보호물질의 종류와 농도, cell 농도 및 건조시간에 대해 실험한 data를 반응표면분석법을 활용하여 분석한 결과, 추정된 최대치의 생존도는 39.3이었으며, 이 최적 조건은 sucrose 11.6(v/v), cell 농도 $1.16{\times}10^{11}$(CFU/ml), 건조 시간 6.18hrs로 나타났다. N.mdeiterranei NCK-4를 동결건조하여 rehydration한 후 계대배양없이 starter cell로서 발효생산에 사용가능성을 검토한 결과, 전배양시 대조군인 FVM(frozen vegetative mycelium)과 거의 비슷하게 cell의 농도를 맞추어 발효함으로써 가능하다. 따라서 이 starter cell은 lyophilization 후 약 18개월간 저장하여 안정하게 사용할 수 있었다.