• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.231-238
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• 1999

The objective of this study was to examine the effect of developmental stage and embryo age of in vitro produced bovine blastocysts after vitrification and thawing. In vitro cultured day 8 blastocysts after IVF were equilibrated 20% ethylene glycol (EG) for 3 min. and were vitrified using EFS40, which is consisted of 40% EG, 18% ficoll, 0.3M sucrose and 10% FBS added in mDPBS for 30 sec. before being plunged into $LN_2$. Also, survival in vitro was assessed by re-expansion and hatching or hatched at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) When the embryos were cultured for 8 day after IVF, 41.0% of the cleaved embryos developed to the blastocysts (early; 7.6%, expanded; 22.9%, hatching; 4.6% and hatched; 5.9%). 2) When the embryos were exposed or vitrified to the freezing solution, the re-expansion of vitrified embryos (73.3%) was significantly lower than that of control and exposed embryos (100, 97.0%) (p<0.05). But the formation rate of hatching or hatched blastocysts of vitrified embryos (66.7, 46.7%) at 48h after thawing was similar to that of exposed embryos (66.7, 39.4%) but not control (100, 100%) (p<0.01). However, in the total cell numbers of those developed hatched blastocysts, there were not significantly different among the treatment groups. 3) When the embryo survival rates by different developmental stage were examined, the re-expansion was not different among the groups $(64.5{\sim}75.6%)$. After warming 48 h, the hatching and hatched formation of early blastocysts (25.8, 9.7%) was significantly lower than those of expanded (69.7, 39.4%) and hatching blastocysts (53.3, 43.3%) (p<0.05). 4) In addition, when the expanded blastocysts at day 7, 8 and 9 were vitrified, the re-expansion of day 8 and 9 embryos was significantly lower than that of day 7 (day 7; 93.9%, day 8; 75.8% and day 9; 87.5%) (p<0.05). However, the rates of development to hatched blastocysts were no difference among the groups (day 7; 36.4%, day 8; 36.4% and day 9; 31.3%). These results suggested that in vitro produced expanded or hatching blastocysts can be efficiently cryopreserved by the two-step vitrification method using EFS40.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.743-760
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• 2014

This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.

• Restorative Dentistry and Endodontics
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• v.31 no.3
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• pp.192-202
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• 2006

The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

• The Korean Journal of Blood Transfusion
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• v.23 no.2
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• pp.115-126
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• 2012

Background: Umbilical cord (UC) is a promising source of mesenchymal stromal cells (MSCs). We compared the characteristics of MSCs from cryopreserved UC with those from fresh tissues, and demonstrated the possibility of UC cryopreservation for acquisition of MSCs from cryopreserved UC. Methods: Each UC was sliced into two types ($1{\sim}2mm^3$ vs. 0.5 cm), and cryopreserved in liquid nitrogen using different media (autologous cord blood plasma, aCBP vs. RPMI 1640). A fresh aliquot of $1{\sim}2mm^3$-sized UC was used as control tissue. After one week, the cryopreserved tissues were thawed and cultured. For the 0.5 cm UC, a slicing step into $1{\sim}2mm^3$ was needed. Cell count, viability, proliferative activity, and surface antigens were determined from harvested MSCs. Several growth factors (EGF, IGF-1, PDGF, TGF-${\beta}$, bFGF, and VEGF), were measured from the culture supernatant. Results: Eleven UC were enrolled in the study. Efficiencies of obtaining MSCs were higher in cryopreserved UC using RPMI 1640, compared with use of aCBP; the same result was observed for 0.5 cm sized UC, compared with $1{\sim}2mm^3$ sized UC. No difference in proliferative activity was observed between MSCs from fresh and cryopreserved UC. The amount of growth factors in culture supernatant using RPMI 1640 was larger than that of fresh tissues. Conclusion: We obtained growth factors from the supernatant as well as MSCs from cryopreserved UC. As with a cord blood bank, in the future, cryopreservation of UC for acquisition of both MSCs and growth factors would be possible in a time of need.

• The Korean Journal of Nuclear Medicine Technology
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• v.22 no.2
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• pp.74-78
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• 2018

Purpose $[^{11}C]$acetate has been proved useful in detecting the myocardial oxygen metabolism and various malignancies including prostate cancer, hepatocellular carcinoma, renal cell carcinoma and brain tumors. The purpose of study was to improve the radiosynthesis yield of $[^{11}C]$acetate on a automated radiosynthesis module. Materials and Methods $[^{11}C]$acetate was prepared by carboxylation of grignard reagent, methylmagnesium chloride, with $[^{11}C]$$CO_2$ gas, followed by hydrolysis with 1 mM acetic acid and purification using solid phase extraction cartridges. The effect of the reaction temperature ($0^{\circ}C$, $10^{\circ}C$, $-55^{\circ}C$) and cyclotron beam time (10 min, 15 min, 20 min, 25 min) on the radiosynthesis yield were investigated in the $[^{11}C]$acetate labeling reaction. Results The maximum radiosynthesis yield was obtained at $-10^{\circ}C$ of reaction temperature. The radioactivities of $[^{11}C]$acetate acquired at $-10^{\circ}C$ reaction temperature was 2.4 times higher than those of $[^{11}C]$acetate acquired at $-55^{\circ}C$. Radiosynthesis yield of $[^{11}C]$acetate increased with increasing cyclotron beam time. Conclusion This study shows that radiosynthesis yield of $[^{11}C]$acetate highly dependent on reaction temperature. The best radiosynthesis yield was obtained in reaction of grignard reagent with $[^{11}C]$$CO_2$ at $-10^{\circ}C$. This radiolabeling conditions will be ideal for routine clinical application.

• Korean Chemical Engineering Research
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• v.59 no.3
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• pp.345-358
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• 2021

This study focuses on the development of the liquid air production process that uses LNG (liquefied natural gas) cold energy which usually wasted during the regasification stage. The liquid air can be transported to the LNG exporter, and it can be utilized as the cold source to replace certain amount of refrigerant for the natural gas liquefaction. Therefore, the condition of the liquid air has to satisfy the available pressure of LNG storage tank. To satisfy pressure constraint of the membrane type LNG tank, proposed process is designed to produce liquid air at 1.3bar. In proposed process, the air is precooled by heat exchange with LNG and subcooled by nitrogen refrigeration cycle. When the amount of transported liquid air is as large as the capacity of the LNG carrier, it could be economical in terms of the transportation cost. In addition, larger liquid air can give more cold energy that can be used in natural gas liquefaction plant. To analyze the effect of the liquid air production amount, under the same LNG supply condition, the proposed process is simulated under 3 different air flow rate: 0.50 kg/s, 0.75 kg/s, 1.00 kg/s, correspond to Case1, Case2, and Case3, respectively. Each case was analyzed thermodynamically and economically. It shows a tendency that the more liquid air production, the more energy demanded per same mass of product as Case3 is 0.18kWh higher than Base case. In consequence the production cost per 1 kg liquid air in Case3 was $0.0172 higher. However, as liquid air production increases, the transportation cost per 1 kg liquid air has reduced by $0.0395. In terms of overall cost, Case 3 confirmed that liquid air can be produced and transported with $0.0223 less per kilogram than Base case.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.1-7
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• 2001

This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.13 no.4
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• pp.348-353
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• 2008

Oxygen isotope has not been used actively in water mass studies because of difficulties on the analysis though it has advantages as a water mass tracer. The most popular method to analysis the oxygen isotope ratio in water samples is equilibration method: isotopic equilibrium of water with $CO_2$ at constant temperature. The precision of oxygen isotope analysis using commercial automatic $H_2O/CO_2$ equilibrator is ${\pm}0.1%o$. This value is not sufficient for studies in open ocean. The object of this study is to improve the analytical precision enough to apply open ocean studies by modification of the instrument. When sample gas is transferred by the pressure difference, the fractionation which is preferential transportation of light isotope can be occurred since the long transportation path between the equilibrator and mass spectrometer. And the The biggest source of error during the analysis is long distance and large volume of the pathway of sample gas between. Therefore, liquid nitrogen trap and high vacuum system are introduced to the system. The precisions of 14 time analysis of same seawater sample are ${\pm}0.081%o$ and ${\pm}0.021%o$ by built-in system and by modified system in this study, respectively.

• Journal of Veterinary Clinics
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• v.24 no.4
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• pp.486-492
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• 2007

Seminal plasma(SP) is usually removed from semen that is to be cryopreserved. However, some reports indicate that SP has beneficial effects on spermatozoa during chilling and freezing. The purpose of this study was to determine the effect of SP on sperm survival by adding SP to the extender before cooling and freezing canine spermatozoa. In replicate experiments, ejaculates obtained from four healthy dogs(1-4 years old) of various breeds were pooled, centrifuged at $300{\times}g$ for 10 min at $25^{\circ}C$, and the supernatant of seminal plasma was decanted. Spermatozoa were suspended in egg yolk-Tris(EYT) buffer. The study comprised two experiments: [Exp 1] Sperm were suspended in EYT extender containing either 0, 20, 40, 80 or 100% SP and were slowly cooled to $4^{\circ}C$ for 2h or held at $25^{\circ}C$ as controls. Sperm concentration was adjusted to $2{\times}10^8/ml$. [Exp II] Sperm samples, each of which contained $1{\times}10^8/ml$, were assigned to nine groups to be frozen. In the first four groups, sperm in EYT containing either 20, 40, 80 or 100% SP were cooled to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol and then were frozen. The final concentrations of SP were 10, 20, 40 or 50%. In the other four groups, sperm in EYT alone were first cooled slowly to $4^{\circ}C$, then diluted to contain final concentrations of EYT+0.6M glycerol plus 10, 20, 40 or 50% SP and then were frozen. Spermatozoa, which chilled in EYT alone and diluted to contain final concentrations of EYT+0.6M glycerol without seminal plasma, and then frozen, was regarded as control. Spermatozoa were frozen at $25^{\circ}C/min$ of cooling rate in plastic straws that were suspended above liquid nitrogen and thawed in water at $38^{\circ}C$ for 1 min. Sperm survival was assayed by determining progressive motility and integrity of plasma and acrosome membranes. Progressive motility was determined by microscopic examination at $200{\times}$ magnification. Membrane integrity was assessed by use of a double fluorescent dye, and acrosome integrity by staining sperm with Pisum sativum agglutinin. The results of the first experiment showed that adding SP did not improve motility of spermatozoa compared to those incubated without SP regardless of temperature. The results of the second experiment showed that spermatozoa suspended in EYT+0.6M glycerol containing SP exhibited the higher progressive motility before being frozen(P<0.05). However, frozen-thawed spermatozoa that had suspended in EYT+0.6M glycerol containing SP showed the similar or lower viability(P<0.05). In summary, although seminal plasma did not affect spermatozoa that were chilled in EYT without cryoprotectant(CPA), addition of seminal plasma to EYT containing CPA did significantly improved progressive motility of canine spermatozoa that were chilled.

• Korean Journal of Agricultural Science
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• v.13 no.1
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• pp.82-89
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• 1986

This experiment was carried out to determine the optimum freezing and thawing rates of the hamster embryos. The female hamsters were induced to superovulate by intraperitoneal injections of 30 i.u. PMSG and mated with males of the same strain of 4 days the PMSG injection. They were killed and embryos were flushed from the oviduct and uterine horn on 3 days after mating. Embryos were flushed with a modified Dulbecco's phosphate-buffered saline and equilibrated with 1.5 M-dimethylsulphoxide by a 3-step procedure. The freezing rates of the samples were $1^{\circ}C/min$ from room temperature to $-6^{\circ}C$ and the samples were seeded at $-6^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were $0.3^{\circ}C/min$ from $-6^{\circ}C$ to $-35^{\circ}C$. From $-35^{\circ}C$ to $-70^{\circ}C$, the rates were divided into $0.1^{\circ}C/min$, $1^{\circ}C/min$ and $10^{\circ}C/min$, respectively. At $-70^{\circ}C$ the samples were plunged directly into liquid nitrogen. The samples were thawed at $4^{\circ}C/min$ and $12^{\circ}C/min$ from $-196^{\circ}C$ to $37^{\circ}C$, and for 2 min in $37^{\circ}C$ water bath, respectively. The average numbers of ovulation points and embryos recovered were 35.1 and 27.0 appearing 77.0% recovery rates. Eight cell embryos in the embryos recovered were 24.8. The survival rates of embryos according to the freezing rates were 55.5~67.7% at $0.1^{\circ}C/min$, 58.8~64.9% at $1^{\circ}C/min$ and 40.5~44.7% at $10^{\circ}C/min$, respectively. The survival rates at $10^{\circ}C/min$ were significantly low. The survival rates of embryos according to the thawing rates were 53.5% at $4^{\circ}C/min$, 53.7% at $12^{\circ}C/min$ and 59.1% in $37^{\circ}C$ water bath. The survival rates, in $37^{\circ}C$ water bath were slightly higher, but we did not find any differences among them. In conclusion, the best freezing rates of hamster embryos were $1^{\circ}C/min$ from the room temperature to $-6^{\circ}C/min$, $0.3^{\circ}C/min$ from $-6^{\circ}C/min$ to $-35^{\circ}C$ and $-0.1^{\circ}C/min$ or $1^{\circ}C/min$ from $-35^{\circ}C$ to $-70^{\circ}C$. The hamster embryos thawed for 2 min in $37^{\circ}C$ water bath showed the best survival rates.