• Journal of Embryo Transfer
• /
• v.23 no.1
• /
• pp.19-24
• /
• 2008

The purpose of this study was to compare the efficiency of slow freezing with that of vitrification method for the cryopreservation of human embryos. Human embryos were derived from in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and the mixed solution of propanedial (1.5, 1.0, 0.5M PROH) and sucrose (0.1M), ethylene glycol (7.5, 15%), dimethyl sulfoxide (7.5, 15% DMSO), sucrose (0.5, 1.0M) and SPS (Serum Protein Substitute) was used for a cryoprotectant for slow freezing and vitrification solution, respectively. Rates of recovery after thawing, morphological normality, post-thaw viability, arrest, morphological abnormality and preimplantation development were compared between two protocols. After freezing-thawing, recovery and survial rate of slow freezing was (88.6% and 73.4%), whereas vitrification was (99.2% and 96.2%) (p<0.05). The arrest rate of slow freezing was significantly lower compared with those of vitrification(8.7% vs 29.9%) (p<0.05). Preimplantation development to the 2-cell (83.8% vs 67.7%), 4-cell (69.0% vs 47.2%) and 8-cell (62.4% vs 37.8%) stages 24, 48 and 72 h after thawing, respectively, were higher in the slow freezing than the vitrification. After slow freezing and vitrification of human embryo at 2-8cell stage, the rate of recovery rate, survival rate and partial damage rate were 92.0% vs 100%, 80.4% vs 96.2% and 52.2% vs 19.0%, respectively. And partial damage rate was significantly lower than those of slow freezing method (p<0.05). These results demonstrate that a slow freezing using PROH is more efficient than a vitrification for cryopreserving the human zygotes, although the vitrification yielded better recovery, survival and partial damage of frozen-thawed 2-8 cell stage embryos than slow freezing method.

• Clinical and Experimental Reproductive Medicine
• /
• v.32 no.2
• /
• pp.177-185
• /
• 2005

Objective: This study was conducted to find an optimal condition for the vitrification of mouse morulae and expanded blastocysts. Materials and Methods: Mouse embryos were obtained at 2-cell stage and cultured to morula and expanded blastocyst stage in Human Tubal Fluid (HTF) medium supplemented with 10% Serum Substitute Supplement (SSS). The vitrification solutions used were EFS30, EFS35 and EFS40 that contains 30%, 35% and 40% ethylene glycol, respectively, with 18% ficoll and 0.5 M sucrose diluted in Dulbecco's phosphate-buffered saline (DPBS) medium supplemented with 10% SSS. The vitrification procedure was performed in EFS solution with three steps, followed by thawing in 6 steps with 0.5 M sucrose, and then survival and hatching-hatched rate per embryos recovered were compared among six groups. Results: After 24 h culture in different vitrification and thawing solution, the survival rate of morula embryos was 94.1%, 85.4% and 59.7% for EFS30, EFS35 and EFS40 group, respectively. Hatching rate of morula embryos after 72 h culture was 30.6%, 25% and 11.3% for EFS30, EFS35 and EFS40 group, respectively. The survival rate of expanded blastocyst embryos after 24 h culture was 90.4%, 98.5% and 100% for EFS30, EFS35 and EFS40 group, respectively. Hatching rate of expanded blastocyst embryos after 48 h culture was 46.2%, 57.6% and 64.3% for EFS30, EFS35 and EFS40 group, respectively. Conclusion: The EFS30 solution was the best for vitrification of mouse morulae. The EFS40 solution was the best for vitrification of mouse expanded blastocysts. The mouse expanded blastocyst was better than mouse morula for vitrification of mouse embryos.

• Journal of Conservation Science
• /
• v.34 no.3
• /
• pp.143-155
• /
• 2018

In this study, we investigated the properties and manufacturing methods of soil mural finishing layers fabricated using animal glue, starch adhesive(wheat paste), and Dobak glue. We assessed the workability and weatherproofing properties of the earthen plaster and finishing layers fabricated using concentrations of 3%, 5%, 7% and 10% for each adhesive. The results showed that a mixture using 3% or 5% starch adhesive or 3% Dobak glue was suitable for constructing the finishing layer. For finishing layers made with animal glue, earthen plaster had poor workability. It was dry and easily broken as the concentrations of animal glue increased. However, specimens made with low concentrations of animal glue did not exhibit surface deterioration after a freezing-thawing test. Therefore, animal glue mixtures could possibly be used for constructing finishing layers in concentrations lower than 3%. Mixtures containing starch adhesive produced plasters with good workability. Additionally, starch adhesive enhanced compression strength. However, when starch adhesive was mixed at concentrations above 7%, the surface exhibited roughening and staining in freezing-thawing tests. When Dobak glue was used in mixtures, it enhanced compression strength, but concentrations above 5% produced specimens with surface cracking. For concentrations of 3%, there were no cracks and the specimens were stable after freezing-thawing tests, so concentrations below 3% of Dobak glue are suitable for constructing finishing layers. We expect this study will be useful for restoring the traditional technology of soil mural finishing layers and suggest using adhesives to construct the finishing layer.

• Food Science and Preservation
• /
• v.13 no.6
• /
• pp.691-696
• /
• 2006

This study was carried out to analyze useful component in freeze-dried persimmon flower powder made from six cultivais. The cultivais were Dogunjosang, Kojongsi, Kabjubaeknok, Chalang, Weolhasi and SangjuDungsi. Powder of persimmon flower was prepared by milling after freeze drying at $-70^{\circ}C$. Crude lipid was the highest in Dogunjosang (57.26%). Major free sugars of the persimmon flowers were fructose ($95{\sim}310mg%$), glucose ($75{\sim}281mg%$) and sucrose ($7{\sim}28mg%$). Major organic acids were malic acid (225 mg% in Kabjubaeknok (Jun. 4th)) and tartaric acid (189 mg% in Kabjubaeknok (Jun. 4th)). Predominant free amino acids were hydroxy-L-proline(25.33 mg% in Weolhasi), L-citrulline (58.83 mg% in SangjuDungsi (May 280)) and L-threonine (11.88 mg% in SangjuDungsi (May 280)). Major phenolic compounds in the persimmon flowers were caffeic acid ($1,946{\mu}g/100 g$ in Kabjubaeknok (Jun. 4th)), p-hydioxybenzoic acid($418{\mu}g/100 g$ in SangjuDungsi (May 29th)) and protocatechuic acid($181{\mu}g/100 g$ in Kabjubaeknok(Jun. 1st)). The results suggest that persimmon flowers be potential materials as useful food ingredients.

• Journal of Embryo Transfer
• /
• v.16 no.3
• /
• pp.239-243
• /
• 2001

Selection of oocyte cryopreservation method is a prerequisite factor for developing an effective bank system. Compared with slow freezing method, the vitrification has various advantages such as avoiding intracellular ice crustal formation. In our previous, we attempted to employ a vitrification method using ethylene glycol and an electron microscope grid for cryopreservation of mouse oocytes. However, A high incidence of spindle and chromosome abnormalities was detected in thawed oocytes after vitrification. We examined whether the addition of a cystoskeleton stabilizer Taxol $^{TM}$, to the vitrification solution could promote the post-thawed survival and subsequent development of stored oocytes. More oocytes developed to the 4-cell (44.7% vs. 69.7%), 8-cell (31.8% vs. 64.2%), morula (24.7% vs. 54.3%), and blastocyst (20.3% vs. 49.2%) stages after the addition of Taxol$^{TM}$ to the cryoprotectant than after no addition. 21 and 26 mouse pups were born after transfer of blastocyst derived from oocytes vitrified without and with Taxol. The addition of Taxol to vitrification solution greatly promoted post-thaw preimplantation development of ICR morose oocytes.tes.

• Journal of Embryo Transfer
• /
• v.19 no.1
• /
• pp.27-34
• /
• 2004

This study evaluated the efficiency and compared with different materials of loading vessels for vitrification-plastic/glass, copper grid and nylon. The loading method, vitrification, cryop-reservation and warming method of the oocytes were examined. The loading samples prepared in manual or company-made and sterilized, loaded the COCs selected on each samples and cultured for maturation during 40 hours, and then exposed sequentially to ethylene glycol solution. Thawing method was reversely treated and exposed for warmed oocytes. After oocytes were thawed, fertilized and cultured in vitro for 3-4 hours, rates of development and morphological appearance were examined. The results were as summarized: ㆍOPS from company-made or hand-made of the hematocrit micropipettes, NLS from fishing line and EMG from company-made for EM were used for loading oocytes, respectively. ㆍThe efficiency of freezing method and loading convenience were orderly higher in OPS, NLS and EMG. The optimal capacity per vessel was orderly lowered in NLS, EMG and OPS, respectively. ㆍAfter oocytes were warmed, the recovery rate, morphology and rate of development were orderly higher in OPS, NLS and EMG, respectively. ㆍIn conclusion, OPS has the advantages of achieving a little more survival and preserving results than other two loading methods.

• Journal of Embryo Transfer
• /
• v.15 no.1
• /
• pp.15-21
• /
• 2000

These studies were undertaken to evaluate morphological normality and development competence in vitro of hyman oocytes following vitrificatioin using ethylene glycol and electron microscopic grid. Human immature oocytes retrieved from natural and stimulated cycles was vitrified at 0 or 48 h and 0, 8 to 15 or 24 to 28 h after maturation culture, respectively. In oocytes retrieved from unstimulated cycle, no signifciant differences were found in morphological normality (56 to 63%) and fertilization (31 to 37%) rates between the times of vitrification. In stimulated patients, however, more oocytes were morphologically normal when vitrified at 24 to 28 h than when vitrified at 0 or 8 to 15 h after maturation culture. Regardlesss of the hormonal stimulation, high cleavage rates(83 to 100%) were obtained in all treatment groups but did not differ significantly. Twenty to 43% of cleaved oocytes developed to the blastocyst stage at 6 days after IVF. These results suggest that vitrified oocytes from unstimulated and stimulated cycles could develop to the blastocyst stage, regardless of the stages of vitrification.

• Journal of Animal Science and Technology
• /
• v.47 no.6
• /
• pp.925-936
• /
• 2005

The present study evaluated whether pentoxifylline added to the freezing extender could improve the sperm characteristics and function in canine frozen semen. Also the conception rate following AI with frozen-thawed semen was investigated. The beneficial effects of pentoxifylline supplementation were visible in motility, viability, acrosome reaction, and tail swelling patterns. Especially, highest sperm viability and function were obtained in the forzen semen supplemented with 1mM pentoxifylline. The follicle size measured by ultrasonography was 6.48 mm, 11.52 mm and 8.9 mm on 11, 13 and 15 days after the onset of natural estrus, respectively and ovulation occurred on 13 and 15 days. The pregnancy rates in bitches inseminated with frozen semen on natural and induced estrus were 71.4% and 75.0%, respectively. There was no significant difference between the pregnancy rates in bitches inseminated with frozen semen following natural and induced estrus, but the litter size was slightly increased in natural cycle.

• Culinary science and hospitality research
• /
• v.20 no.1
• /
• pp.178-188
• /
• 2014

Two kinds of anticaking agents (dextrin, polydextrose) were combined with kiwifruit paste at 5% w/w ratio and freeze-dried to prepare a powdered material. The physiochemical characteristics of kiwifruit powders with anticaking agents were compared with those without anticaking agents as the control. The yield was higher in the powders with anticaking agents than the control. Moisture content, acidity, and total phenolics were lower in the powders with anticaking agents than the control. The contents of vitamin C was higher in the powders with anticaking agents than the control, but was no significant difference with different anticaking agent types. There were no significant differences in free sugar content (fructose, glucose, total sugar) and organic acid content (oxalic acid, lactic acid, total) depending on the anticaking agent types. Hunter's L-value was significantly high in the order of the samples with dextrin, the control, and polydextrose, while a-value showed an opposite tendency. Browning index, water solubility, and swelling power didn't show any significant difference. However, the hygroscopicities with elapsed time were lower in the powders with anticaking agents than the control. Therefore, the kiwifruit powder combined with dextrin or polydextrose as an anticaking agent at 5% w/w ratio could be used as a food biomaterial with a good quality in moisture, vitamim C, color value, browning index, water solubility, and hygroscopicity.

• Journal of Embryo Transfer
• /
• v.9 no.2
• /
• pp.173-180
• /
• 1994

The present experirnents on cryopreservation were carried out to investigate effect of solution toxicity, equilibration time and cell stages on the post-thaw survival of mouse morulae and blastocyst embryos cryopreserved by vitrification in EFS solution. The mouse embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen and thawed rapidly. After the mouse morulae embryos were exposed to EFS solution for 2 and 5 ruin. at room temperature and then they were washed in 0.5 M sucrose solution and basal mediurn(D-PBS + 10% FCS), they were cultured to examined cryoprotectant toxicity induced injury during exposure, most of embryos developed to expanded blastocysts(100 and 90.0%). However, when the exposure time was extended to 10 and 20 min, these development rates dropped dramatically in 10 ruin. (75.0%) and 20 ruin. (4.5%), respectively. When the compacted morulae were vitrified in EFS solution after equilibration for 2 and 5 min, the embryos have developed to normal blastocyst following thawing, washing and culture processes was 89.3 and 89.6%. However, when the exposure time was expanded to 10 ruin, this survival rate dropped to 68.8%. When the blastocyst were vitrified in EFS solution after equilibration for 2, 5 and 10 minutes, the survival rate of embryos which developed to normal blastocyst following thawing and culture processing were 58.5, 46.7 and 22.4%, respectively. The optimal time of equilibration of mouse morula and blastocysts in EFS solution seemed o be 2 and 5 ruin.