• Reproductive and Developmental Biology
• /
• v.29 no.3
• /
• pp.193-199
• /
• 2005

The cryopreservation of germ cells, sperm and embryos, has been largely used to increase the effect of artificial reproductive techniques for human infertility, but the efficiency of germ cell cryopreservation has been conkoversial till now. Thus, the effect of the cryopreservation of human sperm used for ICSI and the effect of the cryopreservation of embryos produced by ICSI on fertilizatiof development and pregnancy were investigated. Sperm freezing did not affect fertilizatiort development and pregnancy rates. Also, there was no significant difference between ejaculated and testicular sperm in ferclizatiort development and pregnancy. Embryo freezing methods, slow freezing and vitrificatior did not differ each other in viability and pregnncy rates. However, ICSI embryo freezing significantly decreased pregnancy rate compared to fresh embryos freezing (p<0.05). In conclusiof this result suggested that cryopreservation of sperm for ICSI did not affect on the resulted embryo development and pregnancy, but ICSI embryo cryopreservation would significantly inhibit pregnancy.

• Korean Journal of Animal Reproduction
• /
• v.16 no.2
• /
• pp.117-123
• /
• 1992

This Study was carried out ot investigate the effects of concentration and equilibration time of cryoprotective aagents on survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and 0.25M sucrose were slowly freezed(from 2$0^{\circ}C$ to -7$^{\circ}C$/-1$^{\circ}C$/min., from -7$^{\circ}C$ to -35$^{\circ}C$/-0.2$^{\circ}C$/min., from -35$^{\circ}C$ to -38$^{\circ}C$/-0.3$^{\circ}C$/min.) by Cell Freezer and directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rate to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.25M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol＋2.0M propanediol was 80.6, 84.7, 75.0 or 78.8%, respectively. 2. The survival rates of porcine embryos after slow frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol＋2.0M propanediol was 80.9, 82.4, 73.1 or 77.1%, respectively. 3. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0M glycerol, 3.0M DMSO, 2.0M propanediol or 2.0M glycerol＋2.0M propanediol was 65.3, 68.6, 63.2 or 59.9%, respectively. 4. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.50M sucrose added 2.0M glycerol, 3.0M DMSO, 2.0 propanediol or 2.0M glycerol＋2.0M propanediol was 67.5, 62.9, 56.9, or 62.8%, respectively. 5. The higher survival rate of porcine embryos was attained at the short period ofequilibration time(5min.) in the freezing medium added 0.25M sucrose and 3.0 DMSO compared to those of 10 or 20min. equilibration time in the same condition.

• Korean Journal of Ichthyology
• /
• v.23 no.1
• /
• pp.75-79
• /
• 2011

This study was conducted to investigate protocol standardization for cryopreservation spermatozoa of the bar-tailed flathead Platycephalus indicus. The suitability of the cryoprotectants, dimethyl sulphoxide (DMSO), glycerol and methanol were tested against three freezing rates and three thawing temperatures. DMSO and glycerol gave significantly higher motile index and survival rates than methanol. Among the freezing rates, freezing at a height of 2 cm above $LN_2$ surface for $10\;min^{-1}$ gave higher motile index and survival rates. In terms of best thawing temperature, $20^{\circ}C$ obtained the highest motility.

• Journal of Convergence for Information Technology
• /
• v.11 no.1
• /
• pp.128-135
• /
• 2021

The possibility and effectiveness of cryopreservation was determined to assess survival rates and improve stock management of thawed embryos of Manila clam, Ruditapes philippinarum. The ideal freezing rates were designed and tested to allow cryoprotectants to equilibrate across the membrane during freezing. Survival rates ranging from 0 to 64.3% were obtained using a stepwise freezing protocol compared with 82.3% control rates. Embryos of Ruditapes philippinarum were equilibrated in 2 CPAs plus sea water for 10 min at 25℃ and then cooled at -1℃/min from 20℃ to -12℃. Straws containing more than 100 embryos were held at 12℃ for 5 min allowing equilibration after seeding and slowly cooled at 2℃/min. to -35℃ for 30 min for equilibration before quenching in liquid nitrogen. Dimethyl sulfoxide (DMSO) is the best cryoprotectant indicated for embryos of R. philippinarum with a survival rate of 64.3±3.28% in the presence of 2.0 M DMSO.

• Korean Journal of Animal Reproduction
• /
• v.13 no.3
• /
• pp.134-140
• /
• 1989

Studies were conducted to seek reasonable methods of rapid freezing of mouse embryos using liquid nitrogen. The effects of the cryoprotectants concentration and the substitution of raffinose to sucrose in freezing and dilution medium on mouse embryo survival rates were determined using the FDA-test. The summarized results are as follows : 1. When 0.3M of sucrose was added into the freezing and dilution medium, FDA scores of embryos were 1.48(1.5M), 3.81(3.0M) and 4.10(4.5M). Higher FDA scores of embryos were obtained in 3.0M and 4.5M glycerol concentrations (P<0.05). 2. With the addition of 0.3M raffinose to the freezing and dilution medium, FDA scores of embryos did not significantly differ between glycerol levels ; 3.97(1.5M), 4.11(3.0M) and 3.54(4.5M). Higher scores of embryos existed in 3.0M glycerol concentration. 3. Concentration of sucrose or raffinose in freezing and dilution medium affected FDA scores of embryos. When sucrose concerations of 0.3, 0.5 and 1.0M were added to the freezing medium, FDA scores of embryos were 3.12, 2.38 and 0, respectively. However, when the same concentrations of raffinose were added to freezing medium, the FDA scores were 4.21, 2.91 and 0. In both cases, better FDA scores of embryos were attained in 0.3M of sucrose or raffinose (P<0.01).

• Journal of Veterinary Clinics
• /
• v.4 no.2
• /
• pp.449-455
• /
• 1987

Quick freezing of bovine embryos was attempted after they were predehydrated at room temperature. Combined solutions of 2M glycerol or 2M ethylene glycol in the presence of either 0.5 or 1.0M sucrose in phosphated buffered saline+20% calf serum were compared. The quick freezing method in which embryos were directly transferred in liquid nitrogen vapor for 2 minutes at - l70$^{\circ}C$ before being plunged into liquid nitrogen was used. Post-thaw survival rates in 2M glycerol and 2M ethylene glycol were high with 0.5 M (55.6% and 53.3%) versus 1.0M(38.1% and 31.6%) sucrose(P < 0.05). But survival rates with 2M glycerol and 2M ethylene glycol were not significantly different. Transfer thawed embryos frozen with 2M glycerol and 2M ethylene glycol by 0.5M sucrose resulted in birthrates of 40.9% and 40.0%, respectively compared to 26.3% and 27.2%, respectively, for 1.0 M sucrose(p<0.05). This was 56.0% for fresh control.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.2
• /
• pp.129-134
• /
• 1998

The objective of this study was to test whether the developmental capacity of human multi-pronuclear (PN) zygotes after ultra-rapid freezing using EM grid can be maintained. For this experiment, multi-PN zygotes which produced in human IVF program were used as an alternatives of normal 2PN zygotes, and they were separated into 3PN or $\geq4PN$ zygotes to compare their in vitro development and cryoinjury according to PN number. As freezing solution, EFS30 which consisted of 30% ethylene glycol, 18% bcoll, 0.5 M sucrose and 10% FBS added D-PBS was used. The result obtained in this experiment was summarized as follows; When the multi..PN zygotes were ultrarapidly frozen and thawed, the high mean percentages (85.5%) were survived. Also when the cleavage rates between control and freezing group were compared with PN number, there were not significantly different in each group (3PN; 81.3% & 85.4% and $\geq4PN$; 90.0% & 95.7%). When the in vitro development rates after thawing were examined, freezing 3PN group (22.0%) was not differed to control 3PN group (38.5%), although the development result of freezing $\geq4PN$ group (45%) was significantly lower than that of control $\geq4PN$ group (44.4%) (p<0.05). These results demonstrate that developmental capacity of human zygote can be obtained by ultra-rapid freezing method using EM grid and EFS30.

• Journal of Microbiology
• /
• v.39 no.3
• /
• pp.240-243
• /
• 2001

The effect of cryoprotectants and suspending solutions on the preservation of marine heterotophic bacteria was investigated. Six halotolerant and four halophilic bacterial isolates suspended in either distilled water or artificial seawater were preserved in glycerol and dimethylsulfoxide at -70$\^{C}$, respectively. After one year of preservation, the recovery rates on the appropriate agar plates were estimated. The survival rate was found to be dependent on the strain tested, regardless of the preservation conditions tested.

• Horticultural Science & Technology
• /
• v.28 no.6
• /
• pp.990-995
• /
• 2010

This study was conducted to compare differences in fruit quality by harvest time of 'Fuji' apple fruit that was frozen on tree by unusual low temperature in that air temperature was under $-3.5^{\circ}C$ for 7 hours. Fruits were harvested at 1 day before, and 2 days and 6 days after freezing damage, respectively. Harvest's soluble solid contents in all treatments was over $14^{\circ}Bx$. Firmness and titratable acidity of fruit harvested after freezing damage was lower than those of fruit harvested before freezing damage. During cold storage, ethylene production of fruit harvested after freezing damage was higher than that of fruit harvested before freezing damage. The reduction in the level of fruit quality during cold storage of fruit harvested after freezing damage was more serious than that of fruit harvested before freezing damage. The reduction of fruit quality during subsequent ambient temperature for 1 week after cold storage of fruit harvested after freezing damage was higher than that of fruit harvested before freezing damage. In comparison of treatments that were harvested at different times after freezing damage, ethylene production and reduction in the level of fruit quality until 8 weeks of cold storage of fruit harvested at 6 days after freezing damage was lower than that of fruit harvested at 2 days after freezing damage. However, this difference by harvest time after freezing damage disappeared after 8 weeks of cold storage. Incidence of flesh browning was not affected by freezing at air temperature under $-3.5^{\circ}C$ for 7 hours.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.7
• /
• pp.1055-1061
• /
• 2014

Innovative freezing technology is currently applied to preserve foodstuffs for long-term storage. Generally, the quality of frozen food is closely related to the types of freezing and thawing processes. In this study, we characterized the physicochemical properties of onions depending on freezing rate. When onions were frozen at $-40^{\circ}C$, freezing rates were 0.1, 0.5, and $0.7^{\circ}C/min$ depending on air-blast quick freezer mode. Onions were thawed by microwave irradiation at 400 W. Hardness of onion dramatically decreased after freezing and thawing compared with blanched onion. However, the fastest freezing rate did not affect hardness. Thawing loss of onion decreased with a faster freezing rate. For morphological observation, onion frozen at a faster rate showed a smaller ice-crystal size. Vitamin C content decreased upon blanching or freezing, but there was no significant difference according to freezing rate. Although free sugar content also decreased upon blanching and freezing, its highest content was at $0.7^{\circ}C/min$ freezing. Among organic acids, malic acid content was highest at $0.7^{\circ}C/min$ freezing. Based on this study, it could be suggested that a faster freezing rate is effective to improve frozen food quality in accordance with preventing tissue damage or minimizing destruction of nutrients.