• Title/Summary/Keyword: Slow-freezing

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Cryopreservation of Oocytes and Embryos by Vitrification (유리화 방법에 의한 난자와 수정란의 동결보존)

  • Gupta, Mukesh Kumar;Lee, Hoon-Taek
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.267-291
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    • 2010
  • Life can be kept in suspended animation either before fertilization at oocyte stage or after fertilization at different stages of embryonic development for a variety of reasons. It not only has potential applications in fertility preservation and management in human but also has important roles in the preservation and management of animal genetic resources, low-cost international movement of selected genetics, and rapid dissemination of germplasm through assisted reproductive technologies (ART) and genetic engineering. Currently, slow-freezing and vitrification are the two approaches by which oocytes and embryos can be cryopreserved for long-term storage. Both of these methods have their own advantages and disadvantages but allow the cryopreservation of oocytes and embryos with comparable efficiency. Vitrification of oocyte and embryos, although proven successful just 13 years after slow-freezing, is generally considered an emerging technology and appears to slow gain acceptance in both animal and human ART despite having controversial storage and contamination issues. In this manuscript, we discuss the basic techniques of oocyt/embryo cryopreservation and review the current status and recent developments in vitrification.

Changes of Internal Pressure during Freezing, Frozen Storage and Thawing of Meats (식육의 동결, 냉동냉장 및 해동시 내부압력 변화)

  • Jeong, Jin-Woong;Kim, Jong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.682-687
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    • 1999
  • The destruction of tissues by volume increase at food freezing is accepted as one of the factor responsible for quality damage. For this reason, the internal pressure developed in meats were investigated with a pressure transducer during freezing, frozen storage and thawing. Increasement of 6.33% for volume and $942.17\;kg/cm^2$ for density at $-20^{\circ}C$ for beef were shown. In quick and slow freezing of beef, internal pressure reached to highest point after reached to the lowest point at initial of the zone of ice crystal formation. The internal pressure was approximately $8{\sim}10\;psig$ and pressure difference was about 1 psig, which was bigger in immersion freezing than that of still-air freezing. During frozen storage of pork, internal pressure of $1.84{\sim}2.32\;psig$ occurred repeatedly as a function of sample weight at material temperature difference of ${\pm}1^{\circ}C$. The internal pressure during thawing of pork was decreased slowly after rapid increase to the maximum for less than 5min at the beginning of thawing. Internal pressure value at thawing was higher than that at freezing in most cases. Internal pressure of beef with thermal equalized freezing was about $1{\sim}4\;psig$, which was lower than that of non-thermal equalized freezing. Also, freezing time was shortened to $10{\sim}20%$.

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Studies on In Vitro Culture, Freezing and Transfer of Korean Native Cattle Embryos Fertilized In Vitro H. Factors Affecting on Survival Rate of Frozen-Thawed Korean Native Cattle Embryos Fertilized In Vitro (한우 체외수정란의 체외배양, 동결보존 및 이식에 관한 연구 II. 한우 체외수정란의 동결 및 융해 후 생존율에 영향을 미치는 요인)

  • 김일화;손동수;이호준;최선호;양병철;이광원;장인호
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.125-135
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    • 1996
  • The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.

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Effects of Cryopreservation of Sperm and Embryos on fertilization, Development and Pregnancy in Int Application (정자와 수정란의 동결이 ICSI 시술에서 수정, 발생 및 임신에 미치는 영향)

  • Min Sung-Hun;Park Yong-Soo;Park Young-Sok
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.193-199
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    • 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.

Effect of Freezing Temperature on the Rehydration Properties of Freeze-Dried Rice Porridge (동결건조 쌀죽의 재수화 특성에 미치는 동결온도의 영향)

  • Koh, So-mi;Rhim, Jong-Whan;Kim, Jeong-Mok
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.509-512
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    • 2011
  • To study the effect of freezing rate on the quality of freeze-dried rice porridge, freeze-dried rice porridge products were prepared with rice porridge pre-frozen at three different temperatures of -20, -40, and -70$^{\circ}C$. The porridge properties such as microstructure, mechanical properties, textural properties, and rehydration rate were determined. Scanning electron microscopy images indicated that fewer air cells were obtained with a larger size of freeze-dried rice porridge frozen at -20$^{\circ}C$ compared with that frozen at -40 and -70$^{\circ}C$. In contrast, quick frozen products at -70$^{\circ}C$ had more dense texture with higher mechanical strength, whereas slow frozen products exhibited higher rehydration rates than those of quick frozen products. In conclusion, the proper choice of pre-freezing temperature plays a decisive role when preparing freeze-dried rice porridge with optimum quality and convenience.

Quality Changes of Salted Chinese Cabbage and Kimchi during Freezing Storage (절임 배추 및 김치의 동결 저장에 따른 품질변화)

  • Koh, Ha-Young;Lee, Hyun;Yang, Hee-Cheon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.1
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    • pp.62-67
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    • 1993
  • The effect of frozen storage on some physicochemical and sensory quality of salted Chinese cabbage and Kimchi were investigated. The texture of the fresh Chinese cabbage was preserved better by emersion quirk freezing or predrying than by air slow freezing or no predrying while no effect was measured on the salted Chinese cabbage. The salted cabbage had less frozen damages than the fresh one and had the similar texture characteristics of the fermented Kimchi. The frozen Kimchi had the similar overall quality to the unfrozen fermented Kimchi in spite of a little higher chewness values. The color of the salted Chinese cabbage was a little changed to pinkish after 3 months frozen storage but Kimchi was maintained the good quality after 6 months.

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Production of Lyopilized Culture of Lactobacillus acidophilus with Preserving Cell Viability

  • Kang, Moo-Heon;Vibhor Saraswat;Lee, Jeewon;Park, Young-Hoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.36-40
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    • 1999
  • Optimal lyophilization process was developed for manufacturing the dried product of Lactobacillus acidophilus with high cell viability. Three major factors, freezing rate, specific surface area of samples, and stabilizer type and their synergy were shown to play a crucial role in the development of an effective lyophilization process. Finally we found an optimal combination among three process parameters mentioned above; an exceptionally high cell survival percentage of 90% was achieved using the 8.28 cm-1 specific surface area of samples, slow freezing rate, and a stabilizer composition of 4% skim milk +1% glycerol +0.1% calcium chloride.

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A Design of RAID-1 Storage using Hard Disk Drive and Flash Memory Drive (하드 디스크 드라이브와 플래시 메모리 드라이브를 활용한 레이드-1 저장장치의 설계)

  • Byun, Si-Woo
    • Transactions of the Society of Information Storage Systems
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    • v.6 no.2
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    • pp.41-46
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    • 2010
  • Recently, Flash Memory Drives are one of the best media to support portable and desktop computers' storage devices. Their features include non-volatility, low power consumption, and fast access time for read operations, which are sufficient to present flash memories as major database storage components for desktop and server computers. However, we need to improve traditional storage management schemes based on HDD(Hard Disk Drive) and RAID(Redundant Array of Independent Disks) due to the relatively slow or freezing characteristics of write operations of SSDs, as compared to fast read operations. In order to achieve this goal, we propose a new storage management scheme called Hetero-Mirroring based on traditional HDD mirroring scheme. Hetero-Mirroring-based storage management improves RAID-1 operation performance by balancing write-workloads and delaying write operations to avoid SSD freezing.

Comparison of Viability on In Vivo and In Vitro-derived Bovine Embryos in Korean Native Cattle (Hanwoo) (한우 체내.외 유래 소 수정란의 생존성 비교)

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Son, Jun-Kyu;Ko, Yeoung-Kyu;Lee, Poong-Yeon;Cho, In-Cheol;Han, Sang-Hyun;Ko, Moon-Suck;Kim, Hyun-Jong
    • Journal of Embryo Transfer
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    • v.26 no.3
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    • pp.195-199
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    • 2011
  • The objective of this study was to evaluate the efficiency of the conventional slow freezing and vitrification methods for cryopreserving in vivo and in vitro-produced bovine embryos. Morphology of post-thawed embryos was evaluated and normal embryos were used for successive culture for 72 h. In experiment I, In embryo viability, There was no significant differences in blastocyst re-expansion rates were found between in vivo and in vitro embryos(89.6% vs. 81.5%). whereas hatched-BL and total cell number rates was significantly higher (p<0.05) for in vivo-derived embryos (76.9%, 136${\pm}$3.6 vs. 43.4%, 107${\pm}$3.8). In experiment II, There was no significant differences in blastocyst re-expansion and Expansion-BL rates were found between in slow freezing and vitrification methods (91.3% vs. 85.7% and 71.4% vs. 75.0%, respectively). in conclusion, These results suggested that the field application for bovine embryo transfer is in part supported by improvements of technologies in embryo conventional slow freezing and vitrification cryopreservation.

Survival Rate, Developmental Competence and Sex Ratio of Post-thawed Hanwoo Embryo Following Biopsy (한우 수정란의 Biopsy 후 배발달율과 동결-융해후 생존성 및 성비)

  • Cho, S.R.;Choi, S.H.;Kim, H.J.;Choe, C.Y.;Jin, H.J.;Cho, C.Y.;Son, D.S.
    • Journal of Animal Science and Technology
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    • v.49 no.2
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    • pp.287-294
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    • 2007
  • The present study was to conducted the sexing efficiency and accuracy of bovine embryo by LAMP (Loop-mediated isothermal amplification) method, the development of the biopsied embryos into re- reformation and the freezability of these blastocysts by slow-freezing and vitrification. In vivo embryos were superovaluted with gonadotropin(Antorin R-10) for 4 days combined with progestrone releasing intravaginal(CIDR) insertion in Hanwoo donors, and in vitro embryos were used blastocyst embryos at Day 7 or Day 8 after post-insemination in vitro. The biopsy of bovine embryo was carried out in a 80μl drop with Ca2+-Mg2+ free D-PBS and the viability of biopsied embryos were evaluated in IVMD medium at over 12 h culture time in 5% CO2 incubator.For embryo sexing, about five or seven blastomeres were isolated from in vitro and in vivo embryos at blastocysts with microblade. and were then subjected to LAMP. The survivability of biopsied embryos were no difference in the development rate to re-formation of blastocysts between in vivo and in vitro embryos(100% and 90% respectively). The rates of sexed embryos were compared according to two groups, the female rate was lower than that the male in the in vivo and in vitro embryos(46% vs, 54% and 40% vs, 60%, respectively). However, there were no difference in the overall sexing ratio between the two groups. The survivability of frozen-thawed sexed embryos were lower in the in vitro than in vivo embryos in the slow-freezing(Group 1) and vitrification method(Group 2), (41.7% vs. 58.8% and 57.1% vs, 77.8. respectively).