• The Korean Journal of Malacology
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• v.26 no.4
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• pp.255-260
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• 2010

This study was conducted to investigate protocol standardization for spermatozoa cryopreservation of the Scapharca broughtonii (Schrenck). Among the freezing rates, freezing at a height of 2 cm above liquid nitrogen surface for 5 minute gave higher activity and survival rate. Among the various diluents, Ringer's solution was the best for S. broughtonii sperm cryopreservation. The suitability of cryoprotectants dimethylsulfoxide (DMSO), dimethylacetamide (DMA), glycerol and methanol were tested against three freezing rates. DMSO gave significantly higher activity and survival rates than others.

• 대한생식의학회:학술대회논문집
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• 1997.11a
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• pp.52-52
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• 1997

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.400-408
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• 2018

Biodiversity has continued to degrade in the $21^{st}$ century due to global warming occasioned by destruction of the environment around the world.. The Nagoya protocol places Korea in a unique position to effectively develop and protect its domestic genetic resources. Microalgae under study in this research contains large amount of antioxidant substances such as beta carotene and astaxanthin, that can be used as biological resource owing to the large amounts of biomass that can be secured through photosynthesis. However, it is difficult to preserve it since cryopreservation method used for long-term preservation is yet to be developed. A basic study for long term cryopreservation was carried out on Nizschia frustulum and Nitzschia amabilis which belong to marine diatoms. As cryoprotectants (CPAs), glycerol, DMSO, and methanol which penetrate into cells were prepared at 5%, 10%, and 15% concentrations each, in case of methanol, it was tested at concentrations of 5%, 10% and 12% by its nature. Two kinds of microalgae, N. frustulum and N. amabilis, were diluted with $10^2$, $10^3$ and $10^4cells\;ml^{-1}$, respectively. The highest survival rate was shown at12% concentration of methanol, and the figures were $6.94{\pm}0.31%$ in N. frustulum and $8.85{\pm}0.16%$ in N. amabilis. As a result of 3 weeks cultivation of thawed microalgae after freezing, the result is shows that N. frustulum increased about 10 times faster and N. amabilis increased about 12 times the original concentration.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.3
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• pp.203-212
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• 2008

Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

• Korean Journal of Animal Reproduction
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• v.24 no.3
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• pp.225-230
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• 2000

This study was designed to investigate effect of cryoprotectant kinds and cell stages on the viability of bovine embryos cryopreserved by vitrification. The oocytes were collected from ovarian follicles of Korean native cows. The follicular oocytes were cultured in TCM-199 medium containing hormone and 10%(v/v) FCS for 24~48hrs in a incubator with 5% $CO_2$, in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 7~10 hrs with spermatozoa capacitated by preincubation. The vitrification solutions of EFS and EDS were consisted of 40%(v/v) ethylene glycol, 18%(v/v) Ficoll and 0.3M sucrose, and 20%(v/v) ethylene glycol, 16.5%(v/v) DMSO and 0.5M sucrose in TCM-199 medium supplemented with 10% FCS, respectively. The embryos were exposed to EFS or EDS at $25^{\circ}C$ and loaded into OPP straw for 30 sec. The plug end of each straws was heat-sealed and straws was slowly immersed into liquid nitrogen(L$N_2$). The results obtained were summarized as follows : 1 . The rates of cleavage and hatching of embryos frozen with vitrification, rapid and slow freezing methods were 67.5%, 27.5% and 42.5%, 20.0% and 52.5%, 25.0%, respectively And rates of cleavage and hatching of embryos frozen with vitrification method were significantly(p<0.05) higher than those in other methods, and the rates were lower than those in control group(82.5% and 37.5%). 2. The rates of cleavage and hatching of embryos were significantly(p<0.05) different between EFS(47.5% and 22.5%) and EPS(52.5% and 27.5%), and the rates were lower than those in control group(82.5% and 37.5%). 3. After vitrification freezing of bovine embryos at zygote, 2 cell, 8 cell, morulae and blastocyst stage, the rate of cleavage and hatching were 25.0% and 15.0%, 32.5% and 20.0%, 37.5% and 20.0%, 52.5%, 27.5%, 47.5% and 25.0%, respectively. And developmental rates to the expended blastocyst stage of embryos frozen at zygote stage was significantly(p<0.05) lower rather than those in 2, 8-cell and morulae stage.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.132-132
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• 2003

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 1999.10a
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• pp.5-10
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• 1999

수의축산분야에서 개발되어진 생식세포 체외조작기술은 의학 및 생물학을 포함한 생명과학 분야에 광범위하게 이용되고 있다. 특히 생식세포의 생존성을 체외에서 장기간 유지시키는 동결보존 기술은 현재 생식의학분야 첨단의료 기술의 발전 및 효율성 향상에 필수적으로 인식되고 있으며, 이에 대한 다양한 연구결과가 소개되고 있다. 본 강의에서는 동결보존기술의 임상적 의미 및 주요 동결기법의 확립과정, 그리고 이를 이용한 최근의 임상연구결과를 간략히 소개하려 한다.

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

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.118-118
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• 2003

홍조류 김속 해조 5종(모무늬돌김, 방사무늬김, 참김, 긴잎돌김, 잇바디돌김)의 사상체를 2단계 냉각법으로 액체질소 중에서 동결보존을 시행하였다. 시료를 여러 가지 동해보호제에 현탁시킨 후 프로그램 냉각기로 4시간에 걸쳐 -4$0^{\circ}C$까지 천천히 동결시켰다. 일차 완만동결 종결 후 즉시 동결용 튜브를 액체질소 중에 수용하여 급속동결 시켰다. 해동시에는 4$0^{\circ}C$의 항온수조에서 대부분의 얼음 결정을 급격히 해동시킨 후 냉각수내에서 완전히 해동시켰다. 생존률은 김 속 해조에서는 neutral red로 염색하여 산정하였으며 50% 해수에 10% DMSO와 0.5M sorbitol 혼합액을 동해보호제로 사용하였을 때의 생존율이 54.6~70.9%였다.

• 대한생식의학회:학술대회논문집
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• 2005.06a
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• pp.107.2-108
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• 2005