• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.6
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• pp.527-535
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• 2020

The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell Keep^TM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell Keep^TM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell Keep^TM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell Keep^TM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

• Journal of Chest Surgery
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• v.39 no.6 s.263
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• pp.427-433
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• 2006

Background: Current vascular prostheses are considered still inadequate for reconstruction of small-diameter vessels. To evaluate the potential use of xenograft vessels as small diameter arterial grafts, we implanted porcine vessels in goats. The grafts were treated with two different processes, freezing and acellularization, before implantation, and gross inspection as well as microscopic examination followed after a predetermined period. Material and Method: Bilateral porcine carotid arteries were harvested and immediately stored at $-70^{\circ}C$ within tissue preservation solution. One of them was designated as frozen xenograft vessel. The other one was put on acellularization process using NaCl-SDS solution and stored frozen until further use. Grafts were implanted in the place of carotid arteries of the same goat. The grafts have remained implanted for 1, 3, and 6 months in three animals, respectively. Periodic ultrasonographic examinations were performed during the observation period. After explantation, the grafts were analyzed grossly and histologically under light microscope. Result: All animals survived the experimental procedure without problems. Ultrasonographic examinations showed excellent patency of all the grafts during the observation period. Gross examination revealed nonthrombotic, patent lumens with smooth surfaces. Microscopic examinations of the explanted grafts showed cellular reconstruction at the 6-month stage in both grafts. Although more inflammatory responses were observed in the early phase of frozen xenografts, there was no evidence of significant rejection. Conclusion: These findings suggest that porcine xenograft vessels, regardless of pre-implantation processes of acelluarization or freezing, can be acceptably implanted in goats, although short duration of observation in a small number of animals may limit this study.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.207-216
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• 2013

This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• Journal of Korean Medicine Rehabilitation
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• v.26 no.4
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• pp.1-8
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• 2016

Objectives Danggwisu-san (Dangguixu-san) is a herbal prescription frequently used to treat pain or swelling caused by contusion. To determine the expiration period through scientific methodology, stability of Danggwisu-san (Dangguixu-san) water extract, a herbal medicine, was examined under various storage conditions and periods. Methods Danggwisu-san (Dangguixu-san) was stored either at room temperature ($23{\pm}2^{\circ}C$), under a refrigerating condition ($4^{\circ}C$) or under a freezing condition ($-18{\pm}2^{\circ}C$) for 0, 1, 2, 3 or 4 weeks and then freeze-dried. Total phenol and total flavonoid amounts were investigated; contents of amygdalin (Prunus persica), paeoniflorin (Paeonia lactiflora), and glycyrrhizin (Glycyrrhiza uralensis) - the marker compounds of Danggwisu-san (Dangguixu-san) - were also analyzed through high performance liquid chromatography (HPLC). Results No significant change in total phenol and total flavonoid amounts was observed under the indicated storage conditions. Moreover, the contents of marker compounds, i.e. amygdalin, paeoniflorin, and glycyrrhizin, did not alter significantly under the indicated conditions, as well. Conclusions Danggwisu-san (Dangguixu-san) was found to be stable up until 4 weeks under the indicated conditions. Further studies on efficacy and long-term stability are warranted to establish the expiration period of Danggwisu-san (Dangguixu-san).

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.2
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• pp.246-253
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• 2012

In this study, we determined the optimum blanching conditions for pretreatment of Aster scaber prior to freezing to ensure its long, safe preservation as a fine cooking ingredient. Frozen-thawed A. scaber did not significantly differ between blanching groups, but the blanched group showed significantly higher Hunter L, a, and b values than the un-blanched group (p<0.05). Higher temperatures and longer treatment times increased softness; hardness did not significantly differ between the blanched and un-blanched groups of frozen-thawed A. scaber (p<0.05). Total bacterial counts and the presence of coliforms seemed to decline with blanching treatments, but treatment temperature and time did not influence this reduction. Over 95% of peroxidase activity was inactivated by blanching treatment but increased slightly after thawing. The sensory evaluation of the frozen-thawed A. scaber by test group showed the A. scaber blanched at $90^{\circ}C$ for 3 min to be the most highly preferred (p<0.05).

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.257-264
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• 2004

For safe preservation of Korean Native Pigs (KNP) as an animal genetic resources and a means to maintain the genetic diversity, we performed to investigate the optimal hormone levels for superovulated gilts and establish the cryopresevation methods of embryos. The reults were as follows; 1. The number of ovulated corpus luteums (CL) and follicles were 12.4, 13.6, 30.0 or 23.3 in hCG 500IU and PMSG 500, 750, 1,000IU or hCG 750IU and PMSG 1,000IU respectively. In the case of PMSG 1,000IU와 hCG 500IU, there showed highest number but were no significance among others. The recovery rate of embryos by the ovulated CL were 59.4-79.2％. 2. The morula stage embryos recovered on Day 4 after insemination were significantly higher than Day 5 (P<0.01), but blastocyst stage embryos recovered on Day 5 were sinificantly higher than Day 4 (P<0.05). 3. The survival rate of expanded blastocyst were 23.5％ in conventional freezing with 1.4 M glycerol.

• Korean Journal of Food Science and Technology
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• v.30 no.3
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• pp.615-623
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• 1998

The garlic juices were extracted and concentrated by heating at $90^{\circ}C$, by using a rotary vacuum evaporator at $45^{\circ}C$, or by freezing at $-50^{\circ}C$ until the volume was reduced to 70% of the original volume. The concentrated juices were kept at $4^{\circ}C\;or\;25^{\circ}C$ for 60 days and the changes of bacteria and color were monitored every 10 days. Flavors of the raw garlic juices, concentrated garlic juices, and garlic powder by hot-air drying were analyzed monthly using GC and GC/MS. The CFUs/mL of mesophilic and psychrotrophic bacteria in the garlic juice concentrated at $90^{\circ}C$ were lower about 1 to 2 log cycles than those of other concentrated juices. Hunter L, a, and b values in the raw garlic raw juice and the juice concentrated at $-50^{\circ}C$ significantly increased at the beginning of storage, in particular, when they were stored at $25^{\circ}C$. The garlic juice concentrated at $45^{\circ}C$ showed relatively low L value but the juice concentrated at $90^{\circ}C$ showed high a value. The main peaks in chromatograms of volatile components identified by GC/MS were 3,3'-thiobis-1-propene, di-2-propenyl disulfide, 3-(methylthio)-1-propene in the raw garlic juice and the concentrated juices, and the juices concentrated at $45^{\circ}C\;or\;-50^{\circ}C$ showed relatively high preservation of garlic flavor.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.3
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• pp.143-148
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• 2018

Objective: The favored method of preserving fertility in young female cancer survivors is cryopreservation and autotransplantation of ovarian tissue. Reducing hypoxia until angiogenesis takes place is essential for the survival of transplanted ovarian tissue. The aim of this study was to investigate the role of angiopoietin-1 (Angpt-1), angiopoietin-2 (Angpt-2), and vascular endothelial growth factor (VEGF) in ovarian tissue grafts that were cryopreserved using two methods. Methods: Ovarian tissues harvested from ICR mice were divided into three groups: group I (control), no cryopreservation; group II, vitrification in EFS (ethylene-glycol, ficoll, and sucrose solution)-40; and group III, slow freezing in dimethyl sulfoxide. We extracted mRNA for VEGF, Angpt-1, and Angpt-2 from ovarian tissue 1 week following cryopreservation and again 2 weeks after autotransplantation. We used reverse transcriptase-polymerase chain reaction to quantify the levels of VEGF, Angpt-1, and Angpt-2 in the tissue. Results: Angpt-1 and Angpt-2 expression decreased after cryopreservation in groups II and III. After autotransplantation, Angpt-1 and Angpt-2 expression in ovarian tissue showed different trends. Angpt-1 expression in groups II and III was lower than in group I, but Angpt-2 in groups II and III showed no significant difference from group I. The vitrified ovarian tissues had higher expression of VEGF and Angpt-2 than the slow-frozen ovarian tissues, but the difference was not statistically significant. Conclusion: Our results indicate that Angpt-2 may play an important role in ovarian tissue transplantation after cryopreservation although further studies are needed to understand its exact function.

• Food Science and Preservation
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• v.15 no.5
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• pp.635-641
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• 2008

This study was conducted to investigate the optimal sub-zero temperature for storage of two garlic cultivars, the northern ecotype 'Jaerae' and the southern ecotype 'Daeseo'. These cultivars of garlic bulbs were stored at different temperature (room temperature, $0^{\circ}C$, and $-4^{\circ}C$) conditions after predrying. Southern ecotype garlic showed higher weight loss than northern ecotype garlic. Northern ecotype garlic at $-4^{\circ}C$ exhibited less than 5% of weight loss during 10 month storage. Sprouting rate was higher in southern than in northern ecotype at room temperature storage. The value of injury by disease and insect was similar between southern ecotype and northern ecotype for 5 month storage, and then northern ecotype showed higher value of injury by disease and insect than southern ecotype. Hunter 'b' value of northern ecotype garlic was the lowest at $-4^{\circ}C$ storage. Northern ecotype garlic showed no cold injury at freezing point temperature storage. However, southern ecotype garlic had $0{\sim}5.3%$ cold injury occurrence at $-4^{\circ}C$ storage. Respiration and ethylene production exhibited the highest at room temperature storage, those had the slight increase at $0^{\circ}C$ and $-4^{\circ}C$ storages. Northern ecotype showed higher enzymatic pyruvic acid and fructan contents than those of southern ecotype. Enzymatic pyruvic acid content increased and fructan contents decreased during storage time.

• Korean Journal of Animal Reproduction
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• v.25 no.3
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• pp.269-275
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• 2001

This study was carried out to investigate the general characteristics such as volume, sperm concentration, sperm motility, sperm abnormality on whole semen, RSP-S and RSP-T semen and fractional semen of small size dogs, and the effect of temperature and preservation time and cryoproservation on motility of whole and RSP-S and RSP-T semen. Multiple ejaculates were collected from small dogs by the digital manipulation of penis. 1. The volume per ejaculate semen, sperm of concentration and motility and abnormal sperm rate of 1st fractional semen were 0.65$\pm$0.09 $m\ell$, 4.52$\pm$0.35$\times$10$^{6}$ cells/$m\ell$, 15.64$\pm$3.85% and 5.50$\pm$0.62%. Also, 2nd fractional semen were 1.25$\pm$0.20$m\ell$, 3.35$\pm$0.48$\times$10$^{6}$ cells/$m\ell$, 96.25$\pm$4.65% and 4.24$\pm$0.46%. And 3rd fractional semen were 1.45$\pm$0.21$m\ell$, 3.55$\pm$0.52$\times$10$^{6}$ cells/$m\ell$, 92.82$\pm$4.24% and 4.66 $\pm$0.58%, respectively. 2. The sperm of concentration and motility and abnormal sperm rates of whole, RSP-S and RSP-T semen were 5.45$\pm$0.82$\times$10$^{6}$ cells/$m\ell$, 95.55 $\pm$4.65%, 4.58$\pm$0.45% and 4.82$\pm$0.36$\times$10$^{6}$ cells/$m\ell$, 90.10$\pm$3.42%, 6.48$\pm$0.68% and 4.55$\pm$0.45$\times$10$^{6}$ cells/$m\ell$, 93.25$\pm$3.85%, 4.82$\pm$0.58%, respectively. 3. The motility of whole, RSP-S and RSP-T semen were higher at 4$^{\circ}C$ than at 38$^{\circ}C$. When preservation temperature was at 4$^{\circ}C$, survival rates of RSP-S and RSP-T sperm were 97.54~6.25% at 1~72 hrs, 97.40~5.62% at 1~100 hrs, respectively. 4. The survival rates of slow and rapid frozen 2nd fraction, RSP-S and RSP-T semen were 67.3$\pm$4.45%, 88.8$\pm$4.46% and 46.4$\pm$3.84%, 74.4$\pm$4.20%, respectively. Survival rates was significantly higher in frozen RSP-S and RSP-T semen than that in control group(8.5$\pm$2.12%).