• Title/Summary/Keyword: Thawed

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Effects of Cryoprotectants on In Vitro Development of Vitrified Immature Porcine Oocytes Following ICSI

  • Lee, Bong-Gu;Rhee, Man-Hee;Kim, Sang-Keun
    • Journal of Embryo Transfer
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    • v.23 no.2
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    • pp.77-80
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    • 2008
  • In the present study, effects of concentration of cryoprotectant solutions on the nuclear maturation of vitrified-thawed porcine oocytes were examined. Also, the developmental capacity of vitrified-thawed immature porcine oocytes following ICSI was investigated. Oocytes were cultured in NCSU-23 medium supplemented with 5% FBS at $38^{\circ}C$ in 5% $CO_2$ and air. The in vitro maturation rate of vitrified-thawed oocytes ($24.1{\pm}2.5%$) was lower than that of the control ($46.0{\pm}3.2%$, p<0.05). The in vitro maturation rate of vitrified-thawed oocytes treated with $1.0{\sim}5.0\;ug$ CB + NCSU- 23 medium were $22.2{\pm}3.0%$, $30.7{\pm}3.2$, $46.3{\pm}3.1%$, $38.5{\pm}3.2%$, respectively. The in vitro maturation rate ($46.3{\pm}3.4%$) of the vitrified-thawed oocytes treated with $3.0\;{\mu}g$ CB for 30 min was the highest of all vitrification groups. When the in vitro developmental rates of the vitrified-thawed (with EDS and EDT) oocytes following ICSI were $18.5{\pm}2.5%$, $16.4{\pm}2.1%$, respectively. This results were lower than the control group ($24.0{\pm}2.5%$).

Offsprings Produced by Transcervically Inseminating Frozen-thawed Semen into Uterus of a Estrus-induced Saanen Goat during Non-breeding Season

  • Yong, Hwan-Yul;Kim, Min-Ah;Bae, Bok-Soo;Kim, Seung-Dong;Jo, Shin-Il;Lim, Yang-Mook;Yoo, Mi-Hyun;Ha, Yong-Hee;Oh, Chang-Shik;Kim, Doo-Hee
    • Journal of Embryo Transfer
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    • v.25 no.2
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    • pp.89-92
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    • 2010
  • We report herein the successful results of estrus induction, sperm cryopreservation and kids born by transcervical insemination of frozen-thawed semen in a Saanen goat. Flugestone acetate (FGA: 60 mg) was inserted into vagina for 15 days. The goat was intramuscularly injected with 400 IU PMSG and 200 IU hCG ($PG600^{(R)}$: Intervet, Korea) a day before withdrawal of the FGA sponge. Follicles and corpora lutea were identified on both ovaries by laparoscopy. Artificial insemination was performed 46 hours after removal of FGA sponge. The concentration of frozen-thawed semen was $3.975{\times}10^8/ml$ and 0.5 ml of frozen-thawed semen was transcervically inseminated into uterine body under anesthesia. Three kids, all females, were born 144 days after artificial insemination. This is the first report producing kids by transcervical insemination of frozen-thawed semen in a Saanen goat of which the estrus was induced by FGA sponges, PMSG and hCG during non-breeding season in Korea.

Expression and Localization of Heat Shock Protein 70 in Frozen-thawed IVF and Nuclear Transferred Bovine Embryos

  • Chung, K.S.;Choi, Y.J.;Song, S.J.;Do, J.T.;Yoon, B.S.;Kim, Y.J.;Lee, H.T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.311-320
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    • 2002
  • The objective of this study was to assess the developmental potential in vitro produced embryos frozen-thawed with the various containers, and also examined expression and localization of heat shock protein 70 at these embryos. For the vitrification, 2-cell, 8-cell and blastocyst stage embryos produced by in vitro fertilization (IVF) and nuclear transfer (NT) were exposed the ethylene glycol 5.5 M freezing solution (EC 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop, and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min. and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid and cryo-loop. However, survival rates by straw were relatively lower than other containers. The use of cryo-loop resulted in only survival of nuclear transferred embryos (43.7%). Also, there embryos after IVF or NT were analysed by semi-quantitive reverse transcription-polymerase chain reaction (RT- PCR) methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNh were higher thawed embryos than control embryos. Immunocytochemistry used to localize the hsp 70 protein in embryos. Two and 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some frozen-thawed embryos. However, in the control, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform In distribution. Therefore, this result suggests that the exploiting Hsp 70 in the early embryos may be role for protection of stress condition for increase viability of embryos within IVF, NT and there frozen-thawed embryos.

Possibility of Instrumental Differentiation of Duck Breast Meat with Different Processing and Storage Conditions (가공 및 저장 방법이 다른 오리 가슴육의 기계적 판별)

  • Sung, Sang Hyun;Bae, Young Sik;Oh, Suk Hwan;Lee, Jae Cheong;Kim, Hyun Joo;Jo, Cheorun
    • Food Science of Animal Resources
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    • v.33 no.1
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    • pp.96-102
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    • 2013
  • The possibility of instrumental differentiation of duck breast meat treated with different processing and storage conditions was investigated for industrial application. Duck breast meats, which were 1) refrigerated (fresh) after slaughter, 2) fresh but applied the torching process for the removal of remaining feathers (fresh-torched), and 3) frozen and thawed (frozen-thawed), were prepared and the torrymeter value and other quality factors were assessed. The torrymeter values of both duck breast meat and skin showed the lowest in frozen-thawed sample during the whole storage period. The drip loss of frozen-thawed sample was higher than those of fresh or fresh-torched ones. The number of total aerobic bacteria was lower in fresh-torched than fresh but both were not different from frozen-thawed at day 1 while no difference found thereafter. Sensory score of frozen-thawed sample was the lowest. The correlation analysis among the torrymeter value and quality factors of duck breast meat revealed that the torrymeter value is closely related with the total aerobic bacterial number, lipid oxidation, drip loss, and storage period but not with color. The results indicate that once the duck breast meat was frozen then thawed, drip loss and sensory quality can be affected and the torrymeter value can differentiate frozen-thawed from fresh and freshtorched duck breast meat.

Improved pregnancy rate and sex ratio in fresh/frozen in vivo derived embryo transfer of Hanwoo (Bos taurus coreanae) cows

  • Jihyun Park;Wonyou Lee;Islam M. Saadeldin;Seonggyu Bang;Sanghoon Lee;Junkoo Yi;Jongki Cho
    • Journal of Animal Science and Technology
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    • v.65 no.4
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    • pp.779-791
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    • 2023
  • This study aimed to assess the effects of embryonic developmental stage, quality grade, and fresh or frozen/thawed conditions on the pregnancy rate and sex ratio of live offspring in Hanwoo (Bos taurus coreanae) cows. The quality and developmental stage of in vivo-derived (IVD) transferred embryos were evaluated using the standard criteria of the International Embryo Technology Society. The recipient cows were synchronized using conventional (estradiol benzoate and progesterone) protocols before embryo transfer. Embryos were transferred to 297 cows, and pregnancy was monitored for 60-70 days after embryo transfer. The pregnancy rates of fresh and frozen/thawed embryos were 56.90% and 52.49%, respectively. Pregnancy rates varied according to embryo quality (56.18% for grade 1 vs. 36.67% for grade 2). Pregnancy rates also varied by developmental stage and cryopreservation (67.86% vs. 63.49% for stage 4-1, 64.00% vs. 54.72% for 5-1, and 50.00% vs. 47.83% for 6-1, in fresh embryos vs. frozen/thawed embryos, respectively). For stage 7-1, the pregnancy rates were 72.73% for fresh embryos and 20.00% for frozen/thawed embryos. In 66 fresh embryos, the sex ratio of live offspring was 5:5, whereas it was 4(female):6(male) for frozen/thawed embryos among the 95 frozen/thawed embryos. The miscarriage rate was approximately 3% higher for frozen/thawed embryos than for fresh embryos (18.1% for fresh vs. 21.1% for frozen). Seasonal fertility rates were 33.3% in spring, 55.67% in summer, 52.8% in autumn, 60.0% in winter. The following male-to-female ratios were observed in different seasons: 6.7:3.3 in spring, 4.0:6.0 in summer, 5.5:4.5 in autumn, and 3.3:6.7 in winter. The current data revealed no significant differences in pregnancy rates between fresh and frozen/thawed IVD embryos. However, there was a lower pregnancy rate with advanced-stage frozen/thawed embryos (stage 7-1). The current study provides comprehensive results for the better optimization of embryo transfer in Hanwoo cattle to obtain the desired fertility rate, pregnancy rate, and sex ratio of calves. These results provide important insights into the factors that influence the viability and success of IVD embryo transfer in Hanwoo cows and may have practical applications for improving breeding programs and reducing production costs.

Cryopreservation of Zona Pellucida Removed and Encased Into Alian Bisected Embryo ofthe Mouse Early Embryos (생쥐 초기배의 라화 분할배와 공투명대내 수납 분할배의 동결보존)

  • 윤창현;강대진;민관식;장규태;오석두
    • Korean Journal of Animal Reproduction
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    • v.15 no.2
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    • pp.103-107
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    • 1991
  • This study was carried out to investigate the survival rate in vitro culture after frozen-thawed to used DMSO(dimethyl sulfoxide), glycerol and ethylene glycol of cryoprotective agents at the zona pellucida removed and encased into alien bisected embryo of the mouse early embryos. The results obtained from this study were as follows : 1. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected morula was 46.6%, 35.8% and 27.3%, total or mean were 36.6%, respectively. 2. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected morula was 70.6%, 65.3% and 66.4%, total or mean were 67.4%, respectively. 3. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected blastocysts was 50.4%, 36.7% and 30.4%, total of mean were 39.2%, respectively. 4. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected blastocysts was 71.1%, 66.7% and 63.9%, total or mean were 67.2%, respectively.

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Effect on Conception for Insemination Method of Frozen Semen in Cheju Native Cattle. 1. Effect of motility and fertility after thawing in frozen semen. (제주한우에 있어서 동결정액 수정방법이 수태에 미치는 영향 제1보 동결정액의 융해방법이 정액상태와 수태에 미치는 영향)

  • 김중규;장덕지
    • Korean Journal of Animal Reproduction
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    • v.8 no.2
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    • pp.79-86
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    • 1984
  • This cepseiment was carried out to cerify the effect of thawing methods and preservative temperature on the sperm motility and fertility after thawing semen with plastic straws in fresh and warm water. Sperm motility in vitro stored at room temperature after thawing were conducted by the various storage hours. A field trial after thawing semen with warmed water in straws from Cheju native cows involving 4 technicians and 800 cows first (or second) services gave the following results. The thawing methods of warmed water for one minute in semen motility were considerably higher than that in iced water during 12 hours after thawing semen, however, the sperm survival index of ice-water shwed a better results according as the time passed away, but not significant differences. Preservative temperaure at 5$^{\circ}C$ (iced water) after thawing gave significantly better results than that of thawed at 3$0^{\circ}C$ (warmed water). The N R rate to 175 inseminations with semen thawed at 15-2$0^{\circ}C$ (fresh water) was 82.8%, 80.9% for 610 inseminations thawed in warm water. Conception rate ofthe semen thawed in warm water for 10-60 secs gave no significant difference among storage hours, because the semen used to be inseminated within one hour almost, but in decreased when semen thawed at the period of one minute over.

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Studies on Effects of Kinds and Concentration of Cryoprotectants, Equilibration Time and Thawing Temperature on the Survival Rate of Rapidly Frozen Porcine Embryos (돼지 수정란의 급속동결시 내동제의 종류와 농도, 평형시간 및 융해온도에 다른 생존성에 관한 연구)

  • 오원진;오건봉;박병권;김상근;이규승
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.15-23
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    • 1994
  • This study was carried out to investigate the effects of concentration, kinds of cryoprotectants, equilibration time, optimum thawing temperature on the survival rate of rapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotectants containing sucrose were directly plunged into liquid nitrogen and thawed in 30, 35 or 37$^{\circ}C$ water bath, Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows : 1. The high survival rate of porcine frozen embryos after rapidly thawed in freezing medium was attained 2.0M DMSO, 2.0M glycerol, 2.0M propanediol, 1.5M ethyleneglycol. 2. The high survival rate of porcine frozen embryos after rapidly thawed in freezing medium was obtained using single cryoprotectant(16.6~40.0%) than mixed cryoprotectants(12.5~33.3%). 3. The eqilibration time on the survival rate of rapidly thawed porcine frozen embryos was attained after short period of time(15.0~33.3%) in the freezing medium higher than long period of time(9.10~30.0%). 4. The thawing temperature on the survival rate of rapidly thawed porcine frozen embryos was attained at 3$0^{\circ}C$ of thawing temperature(33.3~40.6%) in the freezing medium higher than 25 or 37$^{\circ}C$ of thawing temperature.

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Effect of Trophoblastic Vesicles Co-Transfer on Pregnancy Rate Following Embryo Transfer in Cattle (소에 있어서 영양아세포구의 공동 이식이 수정란이식 수태율에 미치는 영향)

  • 양보석;오성종;임기순;박성재;양병철;김경남
    • Journal of Embryo Transfer
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    • v.16 no.1
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    • pp.29-34
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    • 2001
  • To investigate the effect of co-transfer of trophoblastic vesicle (TV) with frozen-thawed in vitro Produced (IVP) bovine embryo on pregnancy rate, IVP blastocysts were transferred to synchronized recipients. Elongated blastocysts were recovered at Day 13 to 15, and dissected more than 4 pieces to removed the embryonic disc. Throphoblastic fragments were cultured for 48 hours to make throphoblastic vesicles (TVs). TVs were cryopreserved in ethylene glycol or vitrification solution and frozen-thawed TVs were co-transferred to recipients with frozen-thawed IVP embryos. 1 The recovery rate of elongated blastocyst on Day 13 to 15 was 22.5% (18/80) and the size of recovered elongated blastocysts was 0.2∼5.0mm. 2. Eighteen elongated blastocysts were dissected into 88 pieces and 61.4% of those pieces were formed to TV (54/88) 3. The viability of frozen-thawed TV in ethylene glycol was higher than in vitrified solution (92.8% vs. 68.8%) 4. The pregnancy rate in co-transfer with frozen-thawed TV and IVP blastocyst was better than transfer only IVP blastocysts (50.0% vs. 23.1%).

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Processing Characteristics of Freeze-Dried Pork Powder for Meat Emulsion Gel

  • Lee, Seonmin;Choi, Yun-Sang;Jo, Kyung;Jeong, Hyun Gyung;Yong, Hae In;Kim, Tae-Kyung;Jung, Samooel
    • Food Science of Animal Resources
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    • v.41 no.6
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    • pp.997-1011
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    • 2021
  • The processing characteristics of freeze-dried pork powder as raw meat for comminuted meat products were compared with those of freeze-thawed pork. The tertiary structural properties, oxidation, and solubility of proteins in the freeze-dried pork powder were investigated. In addition, the properties of the emulsion gels manufactured with freeze-dried pork powder (GFD) and freeze-thawed pork (GFT) at 1.5% and 2.0% NaCl were evaluated. The surface hydrophobicity and intrinsic tryptophan fluorescence intensity of myofibrillar proteins between the freeze-dried pork powder and freeze-thawed pork were similar. However, freeze-dried pork powder had higher carbonyl compounds and lower solubility of sarcoplasmic and myofibrillar proteins than freeze-thawed pork (p<0.05). GFD had higher cooking loss than GFT in 2.0% NaCl, and lower hardness and a* value of GFD were observed regardless of NaCl level (p<0.05). Moreover, GFD had higher malondialdehyde content than GFT at the two NaCl concentrations (p<0.05). Therefore, our study demonstrated that freeze-dried pork powder has lower functional properties than freeze-thawed pork as raw meat for comminuted meat products.