• Title/Summary/Keyword: frozen-thawed

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Freezing Methods of Canine Semen to Achieve Good post-Thaw Viability of Sperm (개 정액의 융해후 정자의 생존율 향상을 위한 동결 방법)

  • 지동범;김용준
    • Journal of Veterinary Clinics
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    • v.17 no.2
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    • pp.420-430
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    • 2000
  • These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

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Some Properties and Curing Effect of Drip from Frozen-thawed Pork meat (돼지고기 드립의 몇가지 특성과 염지 효과)

  • 김미숙
    • The Korean Journal of Food And Nutrition
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    • v.12 no.4
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    • pp.370-374
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    • 1999
  • This study was carried out to some properties and curring effect of drip obtained from frozen-thawed park loin ham belly and imported belly by thawing process at 4$^{\circ}C$. Moisture content and pH value of drips were 88.05~90.85% and 5,72~6.05 and do not show significant differences between each samples. Protein contents were 11.07, 8.85, 8,76 and8,13% in the drips from domestic pork loin, ham, belly and imported belly, respectively. Approximately 99% of the drip were constituted with moisture and protein in any part of domestic pork and imported belly. Glutamic acid proline glycine, alanine and lysine were the predominant amino acid in the drips. Curing process of the drip by nitrite increased the pH value and total amino acid content. The residual nitrite decreased during the period of curing and total plate counts in drip with nitrite did not reach 1$\times$105CFU/g until 7 days.

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Influence of freeze-thawed cycles on pork quality

  • Tippala, Tiprawee;Koomkrong, Nunyarat;Kayan, Autchara
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1375-1381
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    • 2021
  • Objective: This study was conducted to determine the effect of freeze-thawed cycles (Fresh meat, F-T 1 cycle and F-T 2 cycles) on the quality characteristics of porcine longissimus dorsi muscle. Methods: A total of 20 three-crossbred pigs (Duroc×[Large White×Landrace]) were randomly obtained from a commercial slaughterhouse in Thailand. Muscle samples were immediately taken from 10 to 11th of the longissimus dorsi for histochemical analysis. The muscles were cut into 2.54 cm-thick chops. A minimum of 20 chops were used for each treatment (fresh meat, freeze-thawed 1 and 2 cycles). Individually chops were packaged in polyethylene bags and frozen at -20℃ for 6 months followed by thawing in refrigerator at 4℃ for 24 h (the 1st freeze-thawed cycle). The freeze-thawed procedure was repeated for two cycles (the 2nd freeze-thawed cycle). Thawing loss, shear force value, citrate synthase activity and muscle fiber characteristics were determined on the muscles. Results: Results showed that increasing of freeze-thawed cycle increased the thawing loss (p<0.01) and citrate synthase activity (p<0.001). Shear force value of fresh meat was higher than freeze-thawed 1 and 2 cycles (F-T 1 cycle and F-T 2 cycles). Freeze-thawed cycles affected muscle characteristics. Muscle fiber area and muscle fiber diameter decreased with an increasing number of freeze-thawed cycles (p<0.001), while the thickness of endomysium and perimysium were increased (p<0.001). Conclusion: Repeated freeze-thawed cycles degraded muscle fiber structure and deteriorated pork quality.

Simplified Slow Freezing Program Established for Effective Banking of Embryonic Stem Cells

  • Kim, Gil Ah;Lee, Seung Tae;Lee, Eun Ju;Choi, Jung Kyu;Lim, Jeong Mook
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.3
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    • pp.343-349
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    • 2009
  • This study was designed to simplify a cryopreservation program for embryonic stem cells (ESCs) by selection of cooling method and cryoprotectant. Commercially available mouse E14 embryonic stem cells (ESCs) were cryopreserved with various protocols, and morphology and viability of the frozen-thawed ESCs and their reactive oxygen species (ROS) production were subsequently monitored. Post-thaw colony-formation of ESCs was detected only after a slow freezing using dimethyl sulfoxide (DMSO) by stepwise placement of a freezing container into a $-80^{\circ}C$ deep freezer and subsequently into -$196^{\circ}C$ liquid nitrogen, while no proliferation was detected after vitrification. When the simplified protocol was employed, the replacement of DMSO with a mixture of DMSO and ethylene glycol (EG) further improved the post-thaw survival. ROS generation in ESCs frozen-thawed with the optimized protocol was not increased compared with non-frozen ESCs. The use of fresh mouse embryonic fibroblasts as feeder cells for post-thaw subculture did not further increase post-thaw cell viability. In conclusion, a simplified slow-freezing program without employing programmable freezer but using DMSO and EG was developed which maintains cell viability and colony-forming activity of ESCs during post-thaw subculture.

Effects of Ohmic Thawing on the Physicochemical Properties of Frozen Pork

  • Kim, Jee-Yeon;Hong, Geun-Pyo;Park, Sung-Hee;Lee, Sung;Min, Sang-Gi
    • Food Science and Biotechnology
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    • v.15 no.3
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    • pp.374-379
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    • 2006
  • This study was carried out to investigate the physicochemical properties of frozen pork muscle which has been thawed using the ohmic thawing process, and to establish the optimal ohmic power intensity. The samples were frozen at $-40^{\circ}C$ and thawed at 0, 10, 20, 30, and 40 V by ohmic thawing. Increasing ohmic power intensity correlated with increased thawing rates. The relationship between ohmic power intensity and thawing rate can be represented as a polynomial function. The pH value decreased with increasing ohmic power intensity (p<0.05). With regard to color measurement, the $L^*$, a, and b values of thawing at all ohmic power intensities were not significantly different. The water holding capacity showed a peak value of 41.62% with an ohmic thawing intensity of 30 V. Cooking losses were lowest at the lowest ohmic thawing intensity of 10 V. Thiobarbituric acid reactive substance (TBARS) levels with all thawing processes were slightly higher than that of the control (p<0.05). Increasing ohmic power intensity did not tend to change the total volatile basic nitrogen (TVBN) value.

Effect of prior cesarean delivery on the outcomes of intracytoplasmic sperm injection

  • Suzan Atteya Gewida;Mohamed Salah Eldeen Abd Rabbo;Mohammed Abd Elmoety El Samra;Hesham Mahmoud Adel Abdel Moneim
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.1
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    • pp.63-68
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    • 2024
  • Objective: This study was conducted to investigate the impact of previous delivery mode on pregnancy outcomes in patients with secondary infertility after frozen-thawed embryo transfer. Methods: This prospective observational study included 140 patients experiencing secondary infertility. Of these, 70 patients had a previous cesarean delivery (CD), while the remaining 70 patients had a previous normal vaginal delivery (NVD). The primary outcome was the implantation rate. The secondary outcomes included rates of clinical pregnancy, chemical pregnancy, miscarriage, and ectopic pregnancy. Results: The comparison of all fertility outcomes between the two groups revealed no statistically significant differences. The implantation rate was 40.4% in the CD group and 41.7% in the NVD group (p=0.842). The clinical pregnancy rate was 50% in the CD group and 49.3% in the NVD group (p=0.932), while the chemical pregnancy rate was 14.6% in the CD group and 19% in the NVD group (p=0.591). The miscarriage rates in the CD and NVD groups were 20% and 17.6%, respectively (p=0.803). One case of tubal ectopic pregnancy occurred in the NVD group (1.4%). Conclusion: The mode of prior delivery did not significantly impact pregnancy outcomes following frozen-thawed embryo transfer.

Study on the Convenient Freezing Method in Boar Semen

  • 김성곤;장현용;박동헌;박춘근;정희태;김정익;양부근
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.278-278
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    • 2004
  • The purpose of this study was to establish the convenient freezing method for more cheap and simple. Semen quality was evaluated the motility, viability, abnormality, acrosome intactness and membrane integrity. And there were also examined the developmental rates of IVM/IVF embryos using frozen-thawed boar semen in each treatment group. (omitted)

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Development of Effective Cryopreservation Method for Mouse Oocytes (생쥐 난자의 효율적인 냉동보존 방법 확립을 위한 연구)

  • Choi, Su-Jin;Kim, Soo-Kyung;Kim, Ji-Sun;Cho, Jae-Won;Jun, Jin-Hyun;Byun, Hye-Kyung
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.75-81
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    • 2004
  • Objective: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. Methods: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1, 2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-$\beta$-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). Results: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. Conclusion: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.

Adaptation of the Hypoosmotic Swelling Test to Evaluate Membrane Integrity of Boar Spermatozoa

  • Jang, Hyun-Yong;Cheong, Hee-Tae;Hwang, Hwan-Sub;Kim, Jong-Taek;Park, Choon-Keun;Lee, Hak-Kyu;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.121-126
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    • 2007
  • The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at $37^{\circ}C$ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at $37^{\circ}C$, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<005). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated ($0.82{\sim}0.94$) but lipid peroxidation vs other estimated factors was negatively correlated ($- 0.90{\sim}- 0.98$). Among the evaluation methods, motility vs Viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this. study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at $37^{\circ}C$ and HOST can substitute the examination of motility, viability and lipid peroxidation.