• 제목/요약/키워드: Freezing Method

검색결과 692건 처리시간 0.031초

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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    • 제29권3호
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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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    • 제40권4호
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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.

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

  • 백청순;서병희;이재현;이경광
    • Clinical and Experimental Reproductive Medicine
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    • 제16권1호
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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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냉동보존된 생쥐배아를 이용한 정도관리에 관한 연구 (Studies on Quality Control by Frozen-Thaw 2-Cell Mouse Embryos)

  • 한선남;김향미;정혜원;오승은;손영수;유한기;안정자;우복희
    • Clinical and Experimental Reproductive Medicine
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    • 제20권2호
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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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우리나라의 동결심도(凍結深度)에 관한 연구(研究) (A Study on the Depth of Frost Penetration in Korea)

  • 홍원표;김명환
    • 대한토목학회논문집
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    • 제8권2호
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    • pp.147-154
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    • 1988
  • 우리나라의 지반(地盤)은 겨울에는 동결(凍結)하고 온난한 계절에는 융해(融解)하는 계절적(季節的) 동결지반(凍結地盤)이다. 이러한 계절적(季節的) 동결지반(凍結地盤)에서는 동결(凍結)과 융해(融解) 상태의 순환과정을 반복하게 되므로 토목건축구조물(土木建築構造物)을 축조(築造)할 때는 동결(凍結)의 영향을 받는 깊이를 알아 두어야만 한다. 본 논문(論文)에서는 우리나라 전국(全國)의 최대동결심도(最大凍結深度) 분포도(分布圖)가 작성 제시(提示)되며 최대동결심도(最大凍結深度)를 추정할 수 있는 새로운 방법이 모색된다. 본(本) 연구(硏究)의 결과(結果), 최대동결심도(最大凍結深度)는 동결지수(凍結指數)의 평방근과 선형(線形) 관계에 있지 않음이 밝혀졌다. 이러한 동결심도(凍結深度)와 동결지수(凍結指數)의 평방근 상이의 비선형성(非線型性)을 보완하고 보다 현실에 근접한 동결심도(凍結深度) 산정(算定)을 위하여 새로운 경험식(經驗式)이 제안(提案)된다. 이 식에는 동결지수(凍結指數) 이외에 흙의 건조단위중량 및 함수비가 고려되어진다. 또한 이 식은 지금까지 사용되어 오고 있는 기존식(旣存式)과 비교(比較) 검토된다.

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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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    • 제30권4호
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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.

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

  • 김상환;강현아;이명섭;서강석;윤종택
    • 한국수정란이식학회지
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    • 제27권1호
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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.

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

  • 윤소영;이은옥;박영권;전종기;정순용;한정식;정병훈
    • 한국추진공학회:학술대회논문집
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    • 한국추진공학회 2010년도 제35회 추계학술대회논문집
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    • pp.795-796
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    • 2010
  • 바이오매스로부터 합성액체오일을 생산하기 위한 방법의 하나로써 Fischer-Tropsch 합성이 세계적으로 주목을 받고 있다. $C_7-C_{15}$ 파라핀의 수첨이성화 반응은 세탄넘버의 향상과 저점도, 유동점, 및 어는점 등의 저온유동특성의 개선을 위하여 디젤연료의 생산 공정에 적용된다. Fischer-Tropsch 합성으로부터 생산되는 Jet fuel 등의 상업적인 제품들은 낮은 끓는점과 유동점을 개선해야 한다. 본 연구는 합성 오일로부터 bio-jet fuel을 제조하기 위한 수첨이성화 반응용 촉매를 개발하는데 있다. 수첨이성화 반응용 백금/제올라이트 촉매의 특성을 분석하고 모델반응으로써 도데칸의 수첨이성화반응 성능을 회분식반응기에서 조사하였다.

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

  • 이동현;정원석;김홍진;김지원
    • 한국구조물진단유지관리공학회 논문집
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    • 제17권5호
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    • pp.130-135
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    • 2013
  • 결빙 방지 분사 시스템은 강설이 예상되거나 진행 중인 상황에서 신속하게 대응 가능하며, 동절기 결빙으로 인해 발생하는 교통사고와 교통 지정체를 감소시킬 수 있는 장점을 가지고 있다. 하지만 현재 국내에는 결빙 방지 분사 시스템의 설치 기준은 마련되어 있지 않다. 본 연구에서는 선형, 기상조건, 일조조건 등을 다양한 도로 조건을 정량화하여 어떤 구간에 우선적으로 결빙 방지 분사 시스템을 적용하여야 하는지 판단할 수 있는 기준을 제시한다. 이를 위해 국내 지역별 기상 조사, 지형지물에 의해 음지 발생 여부 분석, 선형과 미끄럼 저항성을 고려한 차량 시뮬레이션 등을 수행하고 이론적 검토를 통해 설치 기준 방법을 정립하였고, 또한 이를 반영하여 시스템의 설치를 판단할 수 있는 소프트웨어 (프로토타입)를 개발하였다.

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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    • 제51권1호
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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.