• Clinical and Experimental Reproductive Medicine
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• 제28권1호
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• pp.33-40
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• 2001

Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

• 한국가축번식학회지
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• 제10권2호
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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%.

• Food Science and Biotechnology
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• 제16권4호
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• pp.626-631
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• 2007

Pre-rigor bovine sternomandibularis muscles were frozen at 3 hr postmortem thawed at various temperatures (18, 2, and $-2^{\circ}C$), and then meat quality and sensory properties were compared with those in chilled muscle (control). The meat thawed at $18^{\circ}C$ had lower ultimate pH, water holding capacity, and sensory scores and higher muscle shortening, thaw drip loss, and shear values than those of the other samples. The samples thawed at $-2^{\circ}C$ had significantly lower muscle shortening and higher sensory scores in tenderness and juiciness than those thawed at 18 and $2^{\circ}C$. Muscle shortening, pH, WHC, shear values, and sensory properties were not significantly different between control and sample thawed at $-2^{\circ}C$. By holding at $-2^{\circ}C$, thaw shortening was prevented and tender meat comparable to the chilled meat was obtained. These results indicate that thaw shortening can be largely eliminated if the frozen pre-rigor muscle is thawed at $-2^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• 제30권4호
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• pp.569-575
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• 2017

Objective: The objective of this research was to study the electrical properties and quality of frozen-thawed chicken breast meat and to investigate the relationship between these parameters at different times of frozen storage. Methods: Thawed samples of chicken breast muscles were evaluated after being kept in frozen storage at $-18^{\circ}C$ for different periods of time (1, 2, 3, 4, 5, 6, 7, and 8 months). Results: The results showed that water-holding capacity (WHC) and protein solubility decreased while thiobarbituric acid-reactive substances content increased with increasing storage time. The impedance module of samples decreased during 8-month frozen storage. Pearson correlation coefficients showed that the impedance change ratio (Q value) was significantly (p<0.05) related to pH, color, WHC, lipid oxidation and protein solubility, indicating a good relationship between the electrical properties and qualities of frozen-thawed chicken breast meat. Conclusion: Impedance measurement has a potential to assess the quality of frozen chicken meat combining with quality indices.

• 한국임상수의학회지
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• 제19권1호
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• pp.80-85
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• 2002

This study was performed to investigate the freezomg condition especially focused on extender composition to achieve good post-thaw viability and motility of canine sperm. Semen were collected from 6 male dogs which had been proved to be fertile in the past and were treated for freezing. Equex-STM paste was contained in both the 1st(3%) and the 2nd(7%) diluent and the 2nd diluent was added to the 1st diluent following glycerol equilibration for an hour and a half. To investigate the effect of Equex-STM paste in the extender on post-thaw canine sperm characteristics, the post-thaw viability, motility, and HOS(Hypoosmotic swelling) values were evaluated according to the different composition of extender with or without Equex-STM paste, thawing conditions, and different thawing media added to thawed semen. 1. Canine sperm removed from seminal plasma and frozen )n Sweden extender containing Equex showed higher post-thaw viability, motility, and HOS values than those frozen in the extender containing Equex-STM paste with seminal plasma and those frozen in the extender without Equex and seminal plasma. 2. Canine sperm frozen in Sweden extender containing Equex-STM paste with 5% glycerol showed higher post-thaw viability, motility, and HOS values than those frozen with 3%, 8% glycerol or 5% DMSO. 3. The canine semen frozen in Sweden extender with 5% glycerol and Equex-STM paste showed higher viability, motility, and HOS values when thawed at $70^{\circ}C$ for 8 seconds than when thawed at $37.5^{\circ}C$ for 1 min and at $18-20^{\circ}C$ for 5 min. 4. TFC (tris -fructose-citrate) and PB S (phosphate buffered saline) medium added immediately to thawed canine semen brought better viability, motility, and HOS values for the sperm than those semen added with TGC(tris-glucose-citrate) and no medium. These results indicated that Equex-STM paste in Sweden extender for freezing the canine sperm which were removed from seminal plasma brought good post-thaw viability and motility of canine sperm. Also of the freezing conditions of canine sperm with the same extender containing Equex, the concentration of 5% glycerol, the thawing condition at $70^{\circ}C$ for 8 sec, and TFC and PBS medium added to the thawed semen brought better post-thaw viability and motility of canine sperm than the other conditions used in this study.

• 한국임상수의학회지
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• 제8권1호
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• pp.103-108
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• 1991

It was carried out to investigate the effect of the kind of medium, the concentration of serum added and the temperature of storage on the survival of mouse and bovine embryos preserved In vitro. The survival of frozen-thawed mouse embryos after cooling at 4$^{\circ}C$ was also investigated. It was possible to preserve the embryos of mouse until 4 days and of cattle in a day without significant decrease of the survival rates. The survival rates of frozen-thawed mouse embryos after cooling at 4$^{\circ}C$ over 3 days were below 20%.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2005년도 제5회 발생공학 국제심포지엄 및 학술대회
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• pp.54-54
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• 2005

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2005년도 제5회 발생공학 국제심포지엄 및 학술대회
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• pp.54-54
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• 2005

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.108-108
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• 2004

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.175.2-175.2
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• 2006