• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.9-14
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• 1989

For the cryopreservation of human embryos this study was accomplished as a preliminary experiment. The purpose of this study is to obtain optimal cryoprotectant, addition and dilution method of cryoprotectant and cooling rate for raising survival of frozon and thawed 2-cell mouse embryos. Seeding was done at $-7^{\circ}C$ and the straw contained embryos was plunged at $-30^{\circ}C$ when the slow cooling was ended. Embryos those developed normally to blastocyst after in vitro culture for over 96 hours were regarded as survival ones. The survival was the rate of number of survival embryos against the recovered embryos. The results are followed : 1. The survivals were 6.3, 71.2 and 67.4% respectively, when Glycerol, DMSO and 1,2-Propanediol were used as cryoprotectant. 2. When sucrose was added in freezing solution, the survival was 69.0%. That was higher than the survival of embryos frozen without sucrose in freezing solution. The difference was not significant. 3. Addition and dilution of cryoprotectant by 4 stepwise raised the survival than by direct, but that was not significant. 4. When embryos were frozen by -0.3, -0.5 and $-1^{\circ}C/min$ before plunged into $LN_2$, the survivals were 67.9, 78.0 and 37.0% respectively. The differnce was significant.

• Applied Chemistry for Engineering
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• v.9 no.4
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• pp.516-522
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• 1998

In this study, a water adsorbent was synthesized by radiation grafting of acrylic acid and multifunctional monomers such as 1,2-propanediol dimethacrylate (PDMA) and 1,1,1-trimethylolethane triacrylate (TMETA) onto cellulose and its subsequent treatment with 5% NaOH. Its absorbency on $H_2O$ and 0.9 % NaCl aqueous solution was examined. The highest absorbency on water and on 0.9% NaCl aqueous solution was obtained from the addition of 0.75 vol % PDDMA and of 1.0 vol % TMETA onto acrylic acid solution, respectively. The absorbency of commercial hygienic band on water and NaCl aqueous solution was 21 g/g and 22 g/g, respectively. However, that for acrylic acid-grafted cellulose including TMETA was 298 g/g and 54 g/g, respectively.

• Journal of Forest and Environmental Science
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• v.12 no.1
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• pp.37-44
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• 1996

Two dilignols composed of ${\beta}$-O-4 structure, a important substructure compound in lignin, was synthesized in high yield in a series of the synthetic studies of lignin model compounds. The dimers were identified with $^1H$ and $^{13}C$-NMR and Mass spectroscopy. The important compound of among them, the final synthetic compound [IV].is called 1-(3,4-dimethoxyphenyl)-2-(2'-methoxy-4'-hydroxymethylphenoxy)-propanediol-l,3. This dimeric lignin model compounds should be usefull for the studies of lignin reactions such as pulping, bleaching, pyrolysis, hydrogenolysis, oxidation, reduction, biodegradation, and chemical utilization.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.95-102
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• 1997

The studies on the carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method of bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1$\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentrations of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as devellpmental rate on in vitro culture or FDA-test. The results are smmarized as followes : 1. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was attained 2.0M glycerol, 2.0M DMSO, 1M or 2.0M propanediol. 2. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was obtained single cryoprotectant(6.7~17.4%) than mixed cryoprotectants(6.7~16.7%). 3. In vitro developmental rate of bovine embryos after rapid frozen-thawing in the freezing medium added 0.25M and 0.50M sucrose were higher cleavage rate than those of sucrose concentration of 0.75M and 1.00M. 4. The freezing methods on in vitro developemental rates of bovine embryos was attained slow freezing method(9.70~15.6%) higher than rapid freezing method(9.4~13.3%).

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

• Clinical and Experimental Reproductive Medicine
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• v.20 no.2
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• pp.165-176
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• 1993

These studies were carried out to investigate the optimal freezing protocol for 2 cell mouse embryos and to find the probability of quality control with 2-cell embryos frozen. The embryos showed the best survival by the protocol composed of a freezing solution with the cryoprotectants(1.5M propanediol + 0.1M sucrose), and a 2-steop thawing method(room temperature, 20 sec-37$^{\circ}C$, 20 sec). The developmental ability of frozen-thaw 2-cell embryos did not differ from that of fresh 2-cell embryos in m-KRB medium with 0.4% bovine serum albumin. But development of frozen-thaw embryos was depended on the supplements of the medium. In the albumin-free medium, the developmental rate(rate of blastocysts) was significantly reduced, compared with that in the medium with 0.4% BSA. Also, when frozen-thaw embryos were cultured in the meduim with human fetal cord serum(HCS), the developmental rate of frozen-thaw embryos was sligtly reduced, compared with that of fresh 2-cell embryos. Finally, frozen-thaw 2-cell mouse embryos were more sensitive to the toxic agent of disposable-plastic syringe. Therefore, toxicity of medium could be effectively detected by frozen-thaw 2-cell mouse embryos.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 2010.06a
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• pp.20-20
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• 2010

The effect of excimer laser annealing on the structural and dielectric behaviors of $PbZr_{0.52}Ti_{0.48}O_3$ (PZT) thin films has been investigated. The amorphous PZT thin films were prepared on Pt/Ti/$SiO_2$/Si substrates by a sol-gel method. The PZT precursor was prepared from lead acetate, zirconium acetylacetonate, and titanium isopropoxide. The starting materials were dissolved in n-propanol and 1,3-propanediol. After, the amorphous PZT thin films were laser-annealed (using KrF excimer laser) as a function of the laser energy density and the number of laser pulse. Structural properties of PZT thin films are characterized by using X-ray diffraction (XRD) and scanning electron microscopy (SEM). The dielectric characterization was done on a RT66A test system and a Agilent 4294A impedance analyzer. The PZT thin films show that excimer laser irradiation drastically improved the crystallization and dielectric properties of the PZT thin films, depending on the energy density and the pulse number.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.69-76
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• 1994

The survival and pregnancy rates were compared between non-frozen embryos and cryopreserved embryos at either pronucleate or 2-4 cell stages using the freezing and thawing techniques being identical in both groups were compared with fresh embryos. 496 embryos were frozen with 1, 2-propanediol and sucrose and 117 2-4 cell stages embryos had been thawed and 79.6 and 66.0% of them respectively were survival. Clinical pregnancy rate was 19.2% for embryos frozen at the pronucleate stage and 19.0% for embryos frozen at the 2-4 cell stages while the pregnancy rate of non-frozen embryos was 21.3%. There were no significant difference in the survival and pregnancy rates of embryos frozen at pronucleate and 2-4 cell stages. The current cumulative pregnancy rate per retrieval in all cycles with frozen zygotes is 35.4 %, consid~ erably higher than observed in single transfers of embryos without cryopreservation(21.3%); predicted pregnancy rate after transfer of all frozen embryos is 43.3 %. It is concluded that firstly, the survival and pregnancy rate of cryopreserved embryos at pronucleate or 2-4 cell stages are very similar to those from their fresh embryos and non-frozen embryos and secondly, cryopreservation substantially enhances pregnancy attainment from in vitro fertilization.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.179-186
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• 2003

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.261-268
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• 1998

The present study was to assess the effect of ultrarapid freezing on the development of 1-cell mouse zygote using cryoprotectants, DMSO (dimethyl sulfoxide) or PROH (1,2-propanediol). We investigated the effect of the type and concentration of cryoprotectant, and of the temperature and time of prefreezing equilibration on their capacity to develop to the blastocyst stage in vitro. The concenration, the equilibration temperature, and the exposure time seemed to serve as an important factor in ultrarapid freezing of 1-cell mouse zygotes. In addition to the exposure time and the concentration of cryoprotectant appeared to playa key role in the development of the embryo. In general, the development of the embryo was more effective at $3^{\circ}C$ than $23^{\circ}C$ and 4.5 M than 3 M for 3 to 5 minutes. At $23^{\circ}C$ the development of the embryo was stimulated by DMSO while at $3^{\circ}C$ it was stimulated by PROH. Thus it has been suggested that there exists a correlation between the concentration of cryoprotectants and exposure time in the development of the embryo. In conclusion, we found that for ultrarapid freezing of mouse 1-cell embryos in DMSO, or PROH-based solution, viability shown optimum depending on the cryoprotectant, the concentration of the cryoprotectant and on the temperature and the duration of equilibration.