• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.311-319
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• 2009

Objective: Human embryonic stem cells (hESCs) have the capacity to differentiate into all of the cell types and therefore hold promise for cell therapeutic applications. In order to utilize this important potential of hESCs, enhancement of currently used technologies for handling and manipulating the cells is required. The cryopreservation of hESC colonies was successfully performed using the vitrification and slow freezing-rapid thawing method. However, most of the hESC colonies were showed extremely spontaneous differentiation after freezing and thawing. In this study, we were performed to rapidly collect of early passage hESCs, which was thawed and had high rate of spontaneously differentiation of SNUhES11 cell line. Methods: Four days after plating, partially spontaneously differentiated parts of hESC colony were cut off using finely drawn-out dissecting pipette, which is mechanical separation method. Results: After separating of spontaneously differentiated cells, we observed that removed parts were recovered by undifferentiated cells. Furthermore, mechanical separation method was more efficient for hESCs expansion after thawing when we repeated this method. The recovery rate after removing differentiated parts of hESC colonies were 55.0%, 74.5%, and 71.1% when we have applied this method to three passages. Conclusion: Mechanical separation method is highly effective for rapidly collecting and large volumes of undifferentiated cells after thawing of cryopreserved early passage hESCs.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.241-248
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• 2014

This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4~8 cell embryos in EG were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG showed better development rates of embryos in 2 and 4~8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

• Korean Journal of Animal Reproduction
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• v.10 no.2
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• pp.175-181
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• 1986

These experiments were carried out to clarify the effects various kinds of cryoprotectants which were frequently used in freezing embryos of domestic animals on the survival of frozen-thawed mouse embryos. As cryoprotectant, glycerol, DMSO and methanol were used and the procedures of adding them in medium were practiced by one-step or six-step adding method. Morphologically normal mouse embryos developed to blastocyst by in vitro culture after freezing and thawing were transferred to pseudopregnant recipients by surgical procedures. The results obtained in these experiments were summarized as follows: 1. The survival rates of the frozen-thawed 8-cell embryos, morulas and blastocysts following one-step addition of glycerol were 83.6, 80.3 adn 70.3%, respectively, while following six-step addition of glycerol, 69.2, 56.3 and 66.7% respectively. 2. When glycerol, DMSO and methanol were used as cryoprotectant under the same condition of freezing and thawing, the survival rates of frozen-thawed embryos were 74.0, 76.1 and 37.6%, respectively. 3. The implantation rate of embryos transferred to pseudopregnant recipients after freezing and thawing was 49.2%.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.171-178
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• 1993

The present study was carried out to investigate the developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulse at output voltage of 2.0 kV/cm for $100{\mu}$ sec to induce cell fusion. The recovery rate and developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos was investigated. 1. The recovery rate of nuclear transplanted 2-cell embryos in normal morphology after freezing and thawing was significantly higher in rapid freezing(DMSO 4.5M) than in slow cooling(p<0.01). 2. When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant. In summary, these experiments have proved that rapid freezing method(DMSO 4.5M) is effective in nuclear transplanted 2-cell mouse embryos. If improved micromanipulation techniques and freezing are combined, nuclear transplantation technique will contribute to the improvement of productivity in livestock animals.

• Journal of the Korea institute for structural maintenance and inspection
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• v.17 no.2
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• pp.117-123
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• 2013

As fiber reinforced polymer (FRP) material is appled for a curved structure such as tunnel, FRP material must has a curved shape. Until now, the curved FRP material has been producted by hand-lay-up or filament winding work. It is impossible for mass production of the curved FRP material by these methods. Also, the quality of product by these methods is lower than that by pultrusion method. New pultrusion method and equipment had been developed for production of FRP material with steady curvature. The objective of this study is to evaluate the effect of freezing and thawing on adhesion of cement concrete with coarse-sand coated FRP in repair and reinforcement of cement-concrete structure using curved FRP material.

• International Journal of Highway Engineering
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• v.18 no.1
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• pp.63-72
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• 2016

PURPOSES : The use of roller-compacted concrete pavement (RCCP) is an environmentally friendly method of construction that utilizes the aggregate interlock effect by means of a hydration reaction and roller compacting, demonstrating a superb structural performance with a relatively small unit water content and unit cement content. However, even if an excellent structural performance was secured through a previous study, the verification research on the environmental load and long-term durability was conducted under unsatisfactory conditions. In order to secure longterm durability, the construction of an appropriate internal air-void structure is required. In this study, a method of improving the long-term durability of RCCP will be suggested by analyzing the internal air-void structure and relevant durability of roller-compacted concrete. METHODS : The method of improving the long-term durability involves measurements of the air content, air voids, and air-spacing factor in RCCP that experiences a change in terms of the kind of air-entraining agent and chemical admixture proportions. This test should be conducted on the basis of test criteria such as ASTM C 457, 672, and KS F 2456. RESULTS : Freezing, thawing, and scaling resistance tests of roller compacted concrete without a chemical admixture showed that it was weak. However, as a result of conducting air entraining (AE) with an AE agent, a large amount of air was distributed with a range of 2~3%, and an air void spacing factor ranging from 200 to $300{\mu}m$ (close to $250{\mu}m$) coming from PCA was secured. Accordingly, the freezing and thawing resistance was improved, with a relative dynamic elastic modulus of more than 80%, and the scaling resistance was improved under the appropriate AE agent content rate. CONCLUSIONS : The long-term durability of RCCP has a direct relationship with the air-void spacing factor, and it can be secured only by ensuring the air void spacing factor through air entraining with the inclusion of an AE agent.

• Journal of the Korea institute for structural maintenance and inspection
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• v.17 no.5
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• pp.31-39
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• 2013

In this study, performance of the post-installed anchor system was evaluated with reduced strength of concrete and anchor. One of the post-installed anchors was selected to performance evaluation. Concrete strength was reduced by freeze-thawing test, and the post-installed anchor strength was reduced by corrosion test. The post-installed Anchor was installed in concrete of freezing and thawing and original concrete, and corroded anchor was installed in original concrete only. Anchor diameter and installation depth of the anchor were the variable for each specimen. Performance of post-installed anchor system of each specimen was evaluated by pullout test. Anchor diameter and installation depth of anchor, it may affect the performance of the post-installed anchor system from the experimental test results. Fracture mode of each post-installed anchor system had occurred differently depending on the durability of concrete and anchor. The anchor pullout strength from the experimental test results was used in order to compare with the results of CCD (Concrete Capacity Design) method, and CCD equation was modified. Modified equation was able to predict the anchor pullout strength of post-installed anchor system in Original concrete and freezing and thawing of concrete.

• Korean Journal of Poultry Science
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• v.45 no.4
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• pp.237-244
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• 2018

In this study, to increase the survival rate of frozen/thaw rooster semen, standard protocols of semen thawing procedures were tested by computer-assisted sperm assay (CASA). We tested 4 different thawing protocols for frozen semen, $5^{\circ}C$ for 2 min, $35^{\circ}C$ for 30 s, $54^{\circ}C$ for 13 s, and $70^{\circ}C$ for 7 s. The pooled semen from 5 to 8 Ogye rooster line was diluted in the HS-1 diluent and frozen in 8% methylacetamide (MA) in liquid nitrogen vapors. To determine standard thawing method, straws were plunged into different temperatures and times. The resulting motilities were recorded by the CASA system. The results of this study showed that the best viability of the spermatozoa was shown by exposure at $5^{\circ}C$ for 2 min. Moreover, the longevity test of thawed sperm at $5^{\circ}C$ for 2 min also supported the higher viability under low temperature preservation of $17^{\circ}C$ for 1 hr. Further research is needed to increase the motility of thawed rooster semen for field application. In addition, the in vivo tests for different rooster lines are also needed for the establishment of avian genetic resource bank.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.4
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• pp.287-293
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• 2001

Objective : To investigate the efficacy of high infusion frequency of liquid nitrogen on pregnancy in human embryo after freezing and thawing. Materials and Methods: 150 infertile patients underwent 162 consecutive thawing-ET cycles. In the high infusion frequency group (Group A), 47 patients (50 cycles) underwent cryopreservation with high infusion frequency of liquid nitrogen. In the low infusion frequency group (Group B), 103 patients (112 cycles) underwent cryopreservation with low infusion frequency of liquid nitrogen. We analyzed the clinical characteristics, fertilization rates, development of embryo, good quality embryo ratio, implantation rates, and pregnancy rates between these two groups. Results: There was no difference between the groups with regard to clinical characteristics (mean age, infertility duration, infertility factors, hormone profile), mean number of oocyte retrieval, fertilization rates, and mean embryo number of transfers. The survival rates in group A was 64.9% (228 of 350 embryos), and among the 228 embryos 190 embryos (83.3%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 65 (34.2%), 29 (15.3%), 35 (18.4%), and 37 (19.5%) of grades 1, 2, 3, and above 4, respectively. The survival rates in group B was 63.8% (482 of 755 embryos), and among the 482 embryos 465 embryos (96.5%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 106 (22.8%), 94 (20.2%), 89 (19.1%), and 112 (24.1%) of grades 1, 2, 3, and above 4, respectively. There was no difference in embryo quality change after the freezing-thawing procedure between the groups. Implantation rates (31.1% vs. 34.3%) were not significant. However hCG positive rates in group A (40%) were higher than group B, but not statistically significant. Clinical pregnancy rate (26% vs. 25.9%), on going pregnancy rates (>20 weeks) were not significant (26% vs. 25%). Conclusion: We compared embryo quality change, survival rates, and pregnancy rates between high infusion frequency group and low infusion frequency group and the results were similar between the two groups. Therefore, high infusion frequency of liquid nitrogen for cryopreservation is a worthy method to preserve in human embryos.

• Proceedings of the Korean Institute of Building Construction Conference
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• 2008.11a
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• pp.227-233
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• 2008

Porous concrete has large continuous voids of 20-30 % by volume, and this concrete is attractive as environmental material in Japan i.e. permeable road pavement, river bank protection with vegetation and green roof system which influence thermal environment. It is necessary to confirm the frost resistance when constructing porous concrete structure in cold region. However applicable test method and evaluation criterion of porous concrete has not defined yet. Therefore, the object of this study is to investigate the frost resistance of porous concrete and this investigation attempts to address this concern by comparing 4 kinds of specified freezing and thawing tests methods (JIS A1148 procedure A/B and RILEM CIF/CDF test) in consideration of actual environment. RILEM freeze-thaw tests are different from JIS A1148 freeze-thaw tests, which are widely adopted for evaluating the frost resistance of conventional concrete in Japan, in water absorption, cooling rate, length of freezing and thawing period, and number of freezing and thawing cycles. RILEM CIF test measures internal damage and is primarily applicable for pure frost attack. CDF test is appropriate for freeze-thaw and de-icing salt attack. JIS A1148 procedure A/B showed extremely low frost resistance of porous concrete if the large continuous voids were filled with water and the ice expansion in the large continuous voids set in during cooling. Frost resistance of porous concrete was improved by mixing coarse aggregate (G7) which particle size is smaller and fine aggregate in JIS freezing and thawing tests. RILEM CIF/CDF test showed that freeze-thaw and de-icing resistance of porous concrete was seems to be superior in that of conventional concrete.