• Journal of Animal Reproduction and Biotechnology
• /
• v.37 no.2
• /
• pp.106-112
• /
• 2022

Cryopreservation of porcine ovarian tissue by vitrification method is a promising approach to preserve genetic materials for future use. However, information is not enough and technology still remains in a challenge stage in pig. Therefore, the objective of present study was to determine possibility of vitrification method to cryopreserve porcine ovarian tissue and to confirm an occurrence of cryoinjuries. Briefly, cryoinjuries and apoptosis patterns in vitrified-warmed ovarian tissue were examined by histological evaluation and TUNEL assay respectively. In results, a damaged morphology of oocytes was detected among groups and the rate was significantly (p < 0.05) lower in vitrification group (25.8%) than freezing control group (67.7%), while fresh control group (6.6%) showed significantly (p < 0.05) lower than both groups. In addition, cryoinjury that form a wave pattern of tissues around follicles was found in the frozen control group, but not in the fresh control group as well as in the vitrification group. Apoptotic cells in follicle was observed only in freezing control group while no apoptotic cell was found in both fresh control and vitrification. Similarly, apoptotic patterns of tissues not in follicle were comparable between fresh control and vitrification groups while freezing control group showed increased tendency. Conclusively, it was confirmed that vitrification method has a prevention effect against cryoinjury and this method could be an alternative approach for cryopreservation of genetic material in pigs. Further study is needed to examine the viability of oocytes derived from vitrified-warmed ovarian tissue.

• Korean Journal of Air-Conditioning and Refrigeration Engineering
• /
• v.8 no.4
• /
• pp.451-462
• /
• 1996

In this study, which focuses on ice storage, a fundamental study in cooling and solidification was performed, including the interesting phenomena of density inversion, supercooling and dendritic ice. A numerical study was performed for natural convection and ice formation considering existence of subcooling and dendritic ice were analyzed numerically by using finite difference method and boundary fixing method. In the mesh, the solid fraction was introduced with adding as a term to the energy conservation equation. A flow in the dendrite was modelled as a flow in a porous medium, and the momentum conservation equation was modified to incorporate resistance forces involved in flows through porous media. A numerical solution of the time dependencies of dendrite area and dense ice front was successfully obtained, and the numerical results were good agreement with experimental results. Based on this methodology, a discussion was made of phenomena and characteristics of cooling and freezing processes under various conditions.

• Proceedings of the Korean Institute of Building Construction Conference
• /
• 2011.11a
• /
• pp.33-34
• /
• 2011

In this study, using the double bubble sheet to anti-freezing method in winter the soil embanking Mock up as a part of the development process was carried out. As results, two layers of the double bubble sheet effect 12.6℃~13.8℃ temperature difference of out door temperature that proved superior insulation and thermal performance of the double bubble sheet.

• Korean journal of food and cookery science
• /
• v.13 no.5
• /
• pp.635-638
• /
• 1997

Dried raddish leaves were prepared by using three different pre-treatment methods (shady sun-drying, freezing after blanching, and shady sun-drying after blanching). Then, the retention of minerals in dried raddish leaves was determined. It was shown that the retention of most minerals (Na, K, Fe, Ca, Mg) except P was higher when shady sun-drying method was used. The retention of P was shown to be the lowest when freezing after blanching method was used.

• Journal of the Institute of Electronics Engineers of Korea SD
• /
• v.46 no.5
• /
• pp.15-24
• /
• 2009

This paper proposes an effective low power BIST architecture using the pattern mapping method for 100% fault coverage and the transition freezing method for making high correlative low power patterns. When frozen patterns are applied to a circuit, it begins to find a great number of faults at first. However, patterns have limitations of achieving 100% fault coverage due to random pattern resistant faults. In this paper, those faults are covered by the pattern mapping method using the patterns generated by an ATPG and the useless patterns among frozen patterns. Throughout the scheme, we have reduced an amount of applied patterns and test time compared with the transition freezing method, which leads to low power dissipation.

• Clinical and Experimental Reproductive Medicine
• /
• v.45 no.3
• /
• pp.110-115
• /
• 2018

Objective: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. Methods: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n = 300), a group that underwent Cryotop vitrification (group 2, n = 300), and a group that underwent our in-house freezing method (group 3, n = 300). Results: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p= 0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p= 0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p= 0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable ($88.99{\pm}10.44$, $88.29{\pm}14.79$, and $86.42{\pm}15.23$, respectively; p= 0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p< 0.001). Conclusion: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.3
• /
• pp.343-349
• /
• 2009

This study was designed to simplify a cryopreservation program for embryonic stem cells (ESCs) by selection of cooling method and cryoprotectant. Commercially available mouse E14 embryonic stem cells (ESCs) were cryopreserved with various protocols, and morphology and viability of the frozen-thawed ESCs and their reactive oxygen species (ROS) production were subsequently monitored. Post-thaw colony-formation of ESCs was detected only after a slow freezing using dimethyl sulfoxide (DMSO) by stepwise placement of a freezing container into a $-80^{\circ}C$ deep freezer and subsequently into -$196^{\circ}C$ liquid nitrogen, while no proliferation was detected after vitrification. When the simplified protocol was employed, the replacement of DMSO with a mixture of DMSO and ethylene glycol (EG) further improved the post-thaw survival. ROS generation in ESCs frozen-thawed with the optimized protocol was not increased compared with non-frozen ESCs. The use of fresh mouse embryonic fibroblasts as feeder cells for post-thaw subculture did not further increase post-thaw cell viability. In conclusion, a simplified slow-freezing program without employing programmable freezer but using DMSO and EG was developed which maintains cell viability and colony-forming activity of ESCs during post-thaw subculture.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.59-64
• /
• 2006

This study was conducted to establish a convenient freezing method of boar semen. Boar semen was cooled until $5^{\circ}C$ for 3 hrs using cell freezer and loaded into straws. Semen straws were frozen in different steps in strofoam box filled with $LN_2$. Highest sperm viability (54.0%) was obtained by 1-step freezing(holding at 10 cm height from the surface of $LN_2$ for 10 min). Sperm viability increased by holding at $-102^{\circ}C$ for 10min (74.0%, P<0.05). In thawing regime, sperm viability was significantly higher in $37^{\circ}C$ group than in $52^{\circ}C$ group. The sperm characteristics did not differ between 1-step and 3-step. After IVF using frozen-thawed boar semen, developmental rate of embryos to the morula+blastocyst stage was in 1-step freezing group than that of 3-step freezing group (27.5 vs 14.7%, P<0.05). The result shows that the 1-step freezing with holding at $-102^{\circ}C$ for 10min before plunging into $LN_2$ is a convenient and easy freezing method for boar semen.

• Journal of Embryo Transfer
• /
• v.29 no.4
• /
• pp.397-401
• /
• 2014

The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

• Korean Journal of Medicinal Crop Science
• /
• v.26 no.1
• /
• pp.1-7
• /
• 2018

Background: Rehmanniae radix preparata (RRP) has been used as a traditional medicine and is one of the most important oriental herbal medicines. However, the physical characteristics of RRP are not suitable for use in industry. The present study was under-taken to determined the preparation method of RRP powder and the physicochemical characteristics of RRP powder by milling under different pre-freezing temperatures. Methods and Results: Moisture content, powder yield, particle size, bulk density, compressive stress, extraction yield, and 5-HMF content of PRR powders by milling with pre-freezing temperatures (-20, -40, -60, and $-80^{\circ}C$) were analyzed, and correlation among these factors was determined. Powder yield increased and particle size decreased in a pre-freezing temperature-dependent manner from -20 to $-60^{\circ}C$. The Hausner ratio increased from 1.186 to 1.225 with decreasing temperature from -20 to $-80^{\circ}C$, whereas compressive stress showed the opposite trend. Extraction yield and 5-HMF content were not significantly different between RRP powder. Significant correlations were observed among pre-freezing temperature and physical characteristics (e.g., yield, particle size, Hausner ratio, and compressive stress). Conclusions: These results suggest that the pre-freezing temperature is an important factor affecting the physical characteristics of PRR powder and applicable to the industrial production of RRP powder.