• Title/Summary/Keyword: Freezing Method

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Analysis of Sperm Ability in Specific Pathogen Free Miniature pig for Production of Bio-Organ

  • Kim T. S.;Cao Y.;Cheong H. T.;Yang B. K.;Park C. K.
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.149-154
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    • 2005
  • The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.

Effects of α-Linolenic Acid and Bovine Serum Albumin on Frozen-thawed Boar Sperm Quality during Cryopreservation

  • Lee, Won-Hee;Hwangbo, Yong;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.40 no.4
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    • pp.33-37
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    • 2016
  • This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

Cryopreservation of Mouse 2-Cell Embryos (생쥐 2세포기배의 동결보존)

  • Baik, C.S.;Suh, B.H.;Lee, J.H.;Lee, K.K.
    • 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.

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Studies on Quality Control by Frozen-Thaw 2-Cell Mouse Embryos (냉동보존된 생쥐배아를 이용한 정도관리에 관한 연구)

  • Han, Sun-Nam;Kim, Hyang-Mee;Jung, Hae-Won;Oh, Seung-Eun;Son, Young-Soo;Yu, Han-Ki;Ahn, Jung-Ja;Woo, Bock-Hee
    • 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.

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A Study on the Depth of Frost Penetration in Korea (우리나라의 동결심도(凍結深度)에 관한 연구(研究))

  • Hong, Won Pyo;Kim, Myung Hwan
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.8 no.2
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    • pp.147-154
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    • 1988
  • Korea has the ground which freezes in winter and melts in warmer seasons by turns. Therefore, in designing civil-structures or buildings on such ground, the depth of seasonal frost penetratio must be considered. In this paper, approximate contours of the maximum depth of frost penetration in Korea is presented. It was found that the maximum depth of frost penetration did not have the linear relationship to square root of the freezing index. In order to establish more reliable method to estimate the maximum depth of frost penetration, a new empirical equation is introduced. In the presented equation, the dry unit weight and water content of soil are considered in addition to the freezing index. And the equation is compared with other previous equations used so far.

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Comparison of Different Vehicles on Human Embryonic Stem Cells using Vitrification

  • Lee, Jae-Ho;Kim, Gi-Jin;Kim, Sin-Ae;Lee, Won-Woo;Lee, Hey-Jin;Lee, Dong-Ryul;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.30 no.4
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    • pp.279-285
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    • 2006
  • Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including micro-droplet (MD), open-pulled straw (OPS) and electron microscopic grid (EM-grid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the post-vitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RT-PCR and immunofluorescence assays. The survival rates of the post-vitrified human ES cells using MD, OPS and EM-grid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slow-freezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slow-freezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.

Effect of TES Extender on Sperm Characteristics and Viability of Frozen Semen in Miniature Pig (TES 보존액이 미니돼지 동결 융해 정자의 생존성 및 성상에 미치는 영향)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Lee, Myeong-Seop;Seo, Kang-Suk;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.45-50
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    • 2012
  • The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ($37^{\circ}C$ for 20 sec, 45 sec and $75^{\circ}C$ for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : $60.3{\pm}2.4$, NAI : $58.6{\pm}2.2%$), TLE ($61.3{\pm}2.4$, $62.2{\pm}2.2%$) extender significantly(p<0.05) increased than that in LEY ($50.2{\pm}2.4$, $54.5{\pm}2.2%$) extender thawed at $75^{\circ}C$ for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE ($66.1{\pm}3.2$, $66.2{\pm}1.0%$) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE ($43.3{\pm}0.5%$) while that in LEY ($63.5{\pm}2.3%$) is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

Catalysts for Hydroisomerization of Synthesis-Oil for Bio-jet fuel Production (Bio-jet fuel 제조용 합성원유 수첨이성화 촉매)

  • Yun, So-Young;Lee, Eun-Ok;Park, Young-Kwon;Jeon, Jong-Ki;Jeong, Soon-Yong;Han, Jeong-Sik;Jeong, Byung-Hun
    • Proceedings of the Korean Society of Propulsion Engineers Conference
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    • 2010.11a
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    • pp.795-796
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    • 2010
  • Interest has been increasing worldwide in Fischer-Tropsch synthesis (F-T) as a method of producing synthetic liquid fuels from biomass. Hydroisomerization of $C_7-C_{15}$ paraffins applies to production of diesel fuel with high cetane number and improved cold flow properties, such as viscosity, pour point and freezing point. The commercial products such as fuel jet produced from F-T synthesis should have low freezing and pour points. In this study, our major aim is to develop a catalyst for hydroisomerization of synthesis-oil for bio-jet fuel. Effects of zeolites and platinum loading on hydroisomerization of dodecane were investigated as a model reaction in a batch reactor.

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Study about the Evaluation of Freezing Risk Based Road Surface of Solar Radiation (도로 노면 일조량에 따른 결빙 위험도 평가 방법에 관한 연구)

  • Lee, Dong-Hyun;Jeong, Won-Seok;Kim, Hong-Jin;Kim, Jiwon
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.17 no.5
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    • pp.130-135
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    • 2013
  • Anti-icing system can reduce traffic accidents and congestion by quickly removing the frozen road surface area. there is no decision criteria for determining application of the Anti-icing system in Korea. In this study, we proposed the decision criteria for determining application of the anti-icing system based on weather and road conditions, i.e., geometric and topographic conditions. Regional climate survey, Solar Radiation analysis, and dynamic vehicle simulation considered road geometry was conducted to standardize the installation method of anti-icing system. Also, we have developed a software that can be determined an installation of anti-icing system.

The protective effect of zinc oxide and selenium oxide nanoparticles on the functional parameters of rat sperm during vitrification

  • Nafiseh Tavakolpoor Saleh;Zohreh Hosseinzadeh;Narges Gholami Banadkuki;Maryam Salehi Novin;Sanaz Saljooghi Zaman;Tohid Moradi Gardeshi
    • Clinical and Experimental Reproductive Medicine
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    • v.51 no.1
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    • pp.20-27
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    • 2024
  • Objective: While sperm freezing (cryopreservation) is an effective method for preserving fertility, it can potentially harm the structure and function of sperm due to an increase in the production of reactive oxygen species. This study aimed to assess the impact of zinc oxide nanoparticles (ZnONPs) and selenium oxide nanoparticles (SeONPs) on various sperm functional parameters, including motility, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), acrosome membrane integrity (ACi), and malondialdehyde (MDA) levels. Methods: Semen samples were collected from 20 Albino Wistar rats. These samples were then divided into six groups: fresh, cryopreservation control, and groups supplemented with SeONPs (1, 2, 5 ㎍/mL) and ZnONPs (0.1, 1, 10 ㎍/mL). Results: Statistical analysis revealed that all concentrations of SeONPs increased total motility and progressive reduction of MDA levels compared to the cryopreservation control group (p<0.05). However, supplementation with ZnONPs did not affect these parameters (p>0.05). Conversely, supplements of 1 and 2 ㎍/mL SeONPs and 1 ㎍/mL ZnONPs contributed to the improvement of PMI and ACi (p<0.05). Yet, no significant change was observed in MMP with any concentration of SeONPs and ZnONPs compared to the cryopreservation control group (p>0.05). Conclusion: The findings suggest that optimal concentrations of SeONPs may enhance sperm parameters during the freezing process.