• Journal of Marine Life Science
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• v.1 no.1
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• pp.70-78
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• 2016

Microalgae are of significant importance for future biotechnological applications. Many microalgae banks or laboratories attempt to maintain various microalgae for further research purposes. Cryopreservation has been preferred to reduce a labor-intensive and costly routine sub-culturing. Cryopreservation can also diminish the genetic drift risk. However, cryopreservation as a long term storage of microalgae method are still in developing progress because it cannot be generalized for all microalgae. Microalgae types, cryoprotectant agents (CPAs) types, freezing and thawing methods are the most important factors that should be considered for cryopreservation. In this short review the basic principles and the current advanced of microalgae cryopreservation methods are discussed with a suggested starting parameters for microalgae cryopreservation.

• Journal of the Korea Institute of Building Construction
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• v.5 no.4 s.18
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• pp.115-120
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• 2005

In recent years, polymer-modified mortars using polymer dispersions have been widely used as finish and repair materials in the construction industry because of their excellent properties compares to those of ordinary cement mortar. Especially, the adhesion improvement of ordinary cement mortar and concrete has attracted a great deal of attention from researchers, and several unique and simply applicable techniques for the adhesion improvement have been developed. The purpose of this study is to evaluate the abrasion resistance of polymer-modified mortar according various curing methods. The polymer-modified mortar are prepared with various polymer-cement ratios, and are subjected to three curing methods such as dry rure, standard cure and freezing and thawing cure after two curing methods, and then tested for abrasion. From the test results, the polymer-modified mortars with various polymer-cement ratios have some superior abrasion resistance compared with plain mortar. The abrasion resistance of polymer-modified mortars increase with an increase in the polymer-cement ratio, and is better under water cure than any other curing methods. It is concluded that the abrasion resistance of cement mortar is markedly improved by modifying of polymer dispersion.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.347-353
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• 1997

This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.1-8
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• 2017

Cryopreservation of germ cells from genetically proven animals could be a source of restoration tools from the risk of extinction or disappearance of wanted characteristics. Using frozen semen, the genetic gains of Korean native cattle have been increased greatly for 70 years. The preservation of genetic resources as a form of frozen semen straw has limited availability due to the numbers. To circumvent this weakness of frozen semen, we tested two re-freezing methods with different initial thawing temperatures using frozen Korean proven semen and rare breed semen from albino, black and chikso breeders. It has been known that human sperm could resist to cryo-damages by repeated freeze-thaw cycles, but not for Korean proven bulls number (KPN) or for rare breeds. Total 7 frozen semem from brindled(2), black(1), Korean Albino(2) and KPN(1) bulls were used for our research. After thawing straws under $5^{\circ}C/2min$ or $37^{\circ}C/40sec$ with low temperature water bath and thermo jug, spermatozoa were re-diluted with triladyl diluents after first thawing and re-frozen. Sperm motilities were compared between animals and treated groups after re-thawing. Mean values of motility and viability of refrozen/thawed sperm for expansion of the number of straws were significantly higher in $5^{\circ}C$ than in $37^{\circ}C$ (P < 0.05). However, the activity of viable sperm thawed at $5^{\circ}C$ was significantly decreased before refreezing. It is estimated that re-freezing of frozen semen from rare Korean native cattle is possible with resistant properties of survived spermatozoa.

• Journal of Conservation Science
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• v.34 no.3
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• pp.143-155
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• 2018

In this study, we investigated the properties and manufacturing methods of soil mural finishing layers fabricated using animal glue, starch adhesive(wheat paste), and Dobak glue. We assessed the workability and weatherproofing properties of the earthen plaster and finishing layers fabricated using concentrations of 3%, 5%, 7% and 10% for each adhesive. The results showed that a mixture using 3% or 5% starch adhesive or 3% Dobak glue was suitable for constructing the finishing layer. For finishing layers made with animal glue, earthen plaster had poor workability. It was dry and easily broken as the concentrations of animal glue increased. However, specimens made with low concentrations of animal glue did not exhibit surface deterioration after a freezing-thawing test. Therefore, animal glue mixtures could possibly be used for constructing finishing layers in concentrations lower than 3%. Mixtures containing starch adhesive produced plasters with good workability. Additionally, starch adhesive enhanced compression strength. However, when starch adhesive was mixed at concentrations above 7%, the surface exhibited roughening and staining in freezing-thawing tests. When Dobak glue was used in mixtures, it enhanced compression strength, but concentrations above 5% produced specimens with surface cracking. For concentrations of 3%, there were no cracks and the specimens were stable after freezing-thawing tests, so concentrations below 3% of Dobak glue are suitable for constructing finishing layers. We expect this study will be useful for restoring the traditional technology of soil mural finishing layers and suggest using adhesives to construct the finishing layer.

• Journal of Genetic Medicine
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• v.5 no.1
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• pp.55-60
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• 2008

Purpose : The ability to perform chromosome analysis of cryopreserved cord blood mononuclear cells is important for future retrospective studies. We compared the karyotypes of cryopreserved cells with cells before cryopreservation. Methods : One cord blood (CB) sample was obtained from normal healthy volunteer. Karyotype analysis was performed before cryopreservation. After mononuclear cell separation with Ficoll-Hypaque, the mononuclear cells were cryopreserved by programmed controlled-rate freezer and then transferred into the liquid nitrogen ($-196^{\circ}C$) for 3 days. After rapid thawing, cytogenetic analysis was performed as the same method for each sample by different conditions. The samples were divided by three groups. The first group was no culture before cryopreservation, the second group was 72 hours culture before cryopreservation, but no 24 hours culture after thawing and the third group was 72 hours culture before cryopreservation and 24 hours culture after thawing. Results : The chromosome analysis was successful in the second and third groups of CB sample. Conclusion : The successful result from CB samples may suggest the usefulness of long-term cryopreservation for retrospective study in various clinical settings including hematologic malignancies.

• Clinical and Experimental Reproductive Medicine
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• v.34 no.2
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• pp.117-124
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• 2007

Objective: The aim of this study was to compare the efficacy of slow freezing with vitrification method for cryopreservation of mouse pronuclear stage embryos. Methods: Mouse pronuclear embryos obtained from superovulated mice and classified into 2 groups of slow freezing and vitrification. Slow freezing solution consisted of 1.5 M PROH, 0.1 M sucrose, while vitrification solution consisted of 40% ethylene glycol, 18% Ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline supplemented with 10% SSS. Recovery and survival rates after thawing and development rates to hatching balstocyst stage were compared between two groups. Results: After freezing and thawing, recovery rate of slow freezing group was 93.8%, whereas vitrification group was 66.5% (p<0.01). Survival rate of recovered embryos were similar between two groups as 83.2% in slow freezing and 87.6% in vitrification. Embryo development rates to 2-cell stage after 24 hrs (77.0% vs 59.1%), 4-cell after 48 hrs (72.6% vs 53.3%), blastocyst after 96 hrs (53.1% vs 40.1%) of thawing were significantly higher in vitrification group than those of slow freezing group, respectively. Conclusion: The vitrification method may provide better developmental competence of frozen-thawed embryos than that of slow freezing method for cryopreservation of mouse pronuclear stage embryos.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1596-1603
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• 1999

Dongchimi (Korean-style fermented radish with juice) products were frozen to prevent further acidification and softening of texture by restraining microbial growth and enzyme activity during storage. Dongchimi juice and radish were separated prior to freezing process. Dongchimi radish was frozen at $-20^{\circ}C,\;-70^{\circ}C$ and immersed in liquid nitrogen and dongchimi juice was frozen at $-20^{\circ}C\;and\;-70^{\circ}C$. Frozen dongchimi samples were thawed with ambient temperatures of $4^{\circ}C\;and\;27^{\circ}C$ and with 915 MHz-microwave, respectively. Dongchimi radish immersed in liquid nitrogen and thawed with 915 MHz-microwave showed the highest pectinesterase activity and hardness, and the lowest polygalacturonase activity and color change, indicating that this quick freezing-quick thawing method can be used for the long-term storage of dongchimi products. Dongchimi juice frozen at $-70^{\circ}C$ and thawed with 915 MHz-microwave retained its pH and titrable acidity, and showed a largest reduction in total aerobic count and lactic acid bacteria.

• Journal of the Korean Applied Science and Technology
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• v.39 no.1
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• pp.52-62
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• 2022

The purpose of this study was to find a way to improve the stability and quality of urinalysis by checking the changes in the measurement values of representative clinical chemistry test items according to the repeated freezing and thawing before the urine test and the thawing process. All subjects were 10 healthy males, and the freeze and thaw stability test was performed using their urine samples. In the case of micro-albumin and amylase, there was no statistical significance at 37℃ with time, but at 42℃ and 60℃, there was a statistically significant change in the results with time. There were statistically significant changes in BUN, creatinine, uric acid, and glucose. As a result of long-term stability, after 7 days, glucose mutation increased and amylase decreased at 60℃. In the case of glucose and amylase, there was a statistically significant change in the results over time. To obtain accurate test results, accurate standardization of urinalysis including appropriate collection, storage, and storage methods of urine samples is required and systematic study of conditions for securing stability for each biomaterial is required.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.98-98
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• 2003

Recently, human embryonic stem (hES) cells have become very important resources for ES cell basic research, cell replacement therapy, and other medical applications; thus, efficient cryopreservation methods for these cells are needed. This study examined whether a newly developed minimum volume cooling (MVC) vitrification method, which was tested through cryopreservation of sensitive bovine oocytes, can be used for freezing hES cells. Feeder-free cultured hES cell (MB03) colonies were mechanically dissected into several small clumps following enzymatic treatment. We compared the freezing efficiency of a slow-cooling method using a cryo-module (0.4-0.6C/min, 20-30 clumps/vial) and MVC vitrification using a modified 0.5-ml French mini-straw designated as a MVC straw (>$20,000{\circ}C$/min, 10 clumps/straw) After thawing, in vitro survival of hES cell clumps was higher for MVC-vitrified cells (80.8%, 97/120) than for slow-cooled cells (38.2%, 39/102). Further, the proliferation rate of surviving MVC-vitrified cells was similar to that of control hES cells from 2 weeks after thawing. In addition, vitrified-thawed hES cells demonstrated a normal karyotype, were positively immunostained for surface marker antibodies (AP, SSEA-4 and TRA-1-60) and the Oct-4 antibody, and could differentiate into all three embryonic germ layer cells in vitro. This result demonstrates that hES cell clumps can be successfully cryopreserved by a newly developed MVC vitrification method without loss of human cell characteristics.