• Title/Summary/Keyword: Swim-up

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Reduction of Polyspermy in Porcine in vitro Fertilization by Modified Swim-UP Method

  • Park, C.H.;B.S. Koo;Kim, M.G.;J.I. Yun;H.Y Son;Lee, S.G.;Lee, C.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.110-110
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    • 2003
  • The high incidence of polyspermic fertilization is one of the major causes lowering the overall efficiency of porcine IVF. The common procedure for IVF involves the co-culture of both gametes in the medium drop, which increases sperm concentration and incidence of polyspermy. Therefore, the present study was carried out to increase the efficiency of porcine IVF by reducing polyspermy using a modified swim-up method. This method modifies conventional swim-up washing by placing oocytes directly at the time of washing. Sperm pellet was prepared in the tube and mature oocytes were placed on cell strainer with $70 \mu m$ pore size (Falcon 2350) at the top of the tube. After insemination, the oocytes were stained for examination. Also, the developmental potential of fertilized embryos was measured to evaluate for the feasibility of this method. While having similar penetration rates in both methods ($86.67 \pm 2.36% to 83.33 \pm 1.36%$), there was a significant reduction of polyspermy in modified swim-up method ($17.50 \pm 1.60%$) compare to the control ($44.1 \pm 3.70%$ (p<0.05). Subsequent culture showed higher rate of blastocyst formation in modified swim-up method (20.44$\pm$0.99%) than the control ($15.73 \pm 3.26%$) (P<0.05), even though there was no significant difference. These results suggest that, by controlling the number of spermatozoa reaching the oocytes, porcine oocytes might be protected from polyspermy in vitro. Also, the developmental potential of the fertilized embryos using this method could be improved by increasing the pool of spermatozoa with better quality. Further optimization of the procedure required to implicate this method in routine porcine IVF.

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Quantitative Selection of Capacitated-sperm with BSA, Caffeine, Heparin and Progesterone (BSA, Caffeine, Heparin 및 Progesteron을 이용한 수정능획득정자의 양적 선별)

  • 김경화;여영근;박영식
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.59-67
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    • 1999
  • To make up the medium for quantitative selection of capacitated-sperm through sucrose layer, the effects of BSA, caffeine, heparin and progesterone on sperm swim-up migration and movement were examined. And the results obtained were as follows; 1. BSA of 4mg/ml in bMSS stimulated sperm migration and movement, and attracted capacitated-sperm. 2. Caffeine of 5mM in bMSS containing 4mg/ml BSA stimulated sperm movement and attracted capacitated-sperm. 3. Heparin of 20$\mu\textrm{g}$/ml in bMAA containing both 4mg/ml BSA and 5mM caffeine stimulated movement and capacitation of sperm. 4. Progesterone of 50$\mu\textrm{g}$/ml in bMSS containing all 4mg/ml BSA, 5mM caffeine and 20$\mu\textrm{g}$/ml heparin (BCHP-MSS) attracted capacitated-sperm. 5. Effect of BCHP-MSS on sperm on sperm attraction was not different from effect of 10% follicular fluid solution (FF-MSS) on sperm swim-up separation. In conclusion, bMSS with 4mg/ml BSA, 5mM caffeine, 20$\mu\textrm{g}$/ml heparin and 50$\mu\textrm{g}$/ml progesterone(BCHP-MSS) was a optimal condition for selection of capacitated-sperm through sucrose layer.

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Comparison of the effect of different media on the clinical outcomes of the density-gradient centrifugation/swim-up and swim-up methods

  • Kim, Eun-Kyung;Kim, Eun-Ha;Kim, Eun-Ah;Lee, Kyung-Ah;Shin, Ji-Eun;Kwon, Hwang
    • Clinical and Experimental Reproductive Medicine
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    • v.42 no.1
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    • pp.22-29
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    • 2015
  • Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF. Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123). Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates. Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.

Separation of Follicular Fluid Components Stimulating Sperm Migration with Chromatographic Paper, $=mu$RPC and Superose Columns (Chromatography용 Paper, $\mu$RPC Column 및 Superose Column을 이용한 정자의 이동을 자극하는 난포액 성분의 분리)

  • 박영식
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.301-312
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    • 1998
  • To efficiently separate a protein stimulating sperm swim-up migration and movement from follicular proteins, the effect of paper chromatography and liquid chromatography with reverse phase column and superose column on protein separation was examined. And the results obtained were as follows; 1. The band component that was separated with paper chromatography stimulated sperm migration and movement depending on its additional levels. Especially, band I component significantly increased sperm migration. But, all components of bands 1, 2 and 3 showed lower sperm migration and movement, compared to follicular fluid at the same additional level. 2. Among the components separated from follicular protein of 2~5mm follicles with reverse phase column ($\mu$RPC), components at retention time (RT) of 3.33, 7.00, 13.87, and 16.6A minutes stimulated sperm migration within a limited range. 3. All components separated from follicular protein of 10mm follicles with $\mu$RPC column didn't stimulate sperm migration and movement. 4. Among the components separated from follicular protein of 2~5m follicles with superose column, components at retention volume (RV) of 1.35 and 0.82 ml significantly stimulated sperm migration and movement. In conclusion, protein components stimulating sperm migration and movement were efficiently separated with superose column in Smart system. Especially, components of RV 1.35 and RV0.82 stimulated sperm swim-up separation.

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Oxygen Toxicity: Behavioral Assessment by Swim Test in Mouse Exposed to Hypoxia and Hyperoxia (수영 시험 모델에서 산소 농도에 따른 운동성 변화와 독성 평가)

  • 김동희;강문철;김재일;이근호;김광열;김형건
    • Toxicological Research
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    • v.19 no.2
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    • pp.99-104
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    • 2003
  • This study was aimed to evaluate the effect of oxygen on motor activity and toxicity in male mice. The modified Porsolt forced swim test (FST) was used and the distance and time of movement by mice were analyzed in 15。C water bath for 20 minutes using the automatic Ethovision videotracking system. Analyses were carried out before and after 20 minutes of exposure to 10%-70% concentration of normobaric oxygen. The effects of inspired oxygen tension on the distance and time of movement showed the similar trends, but changes in distance were more prominent. Both the distance and time of movement increased after exposure to 30% and 40% oxygen concentration. The distance and time of movement also increased upon exposure to 50% and 60% oxygen. In contrast, increases En movement and time under exposure to 21% oxygen concentration were suppressed when exposed to over 50% oxygen concentration. With exposure to 10% oxygen, there was a significant decrease in the distance of movement and a slight suppression of movement time. During the swim test, 12.5%, 37.5%, and 87.5% of the mice drowned after exposure to 10%, 60%, and 70% oxygen concentration, respectively. These results suggest that motor activity can be enhanced by inspired oxygen up to 40% concentration. When hypoxic and hyperoxic oxygen exposure over 50%, motor activity is reduced and toxicity may be induced.

Analysis of Sperm Chemoattractant in Follicular Fluid (난포액내 정자유인물질의 분석)

  • 박영식
    • Journal of Embryo Transfer
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    • v.14 no.1
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    • pp.47-57
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    • 1999
  • Among proteins separated from methanol extract of follicular fluid with superose column, the components inducing sperm swim-up separation through sucrose layer were analysed with superose column in Smart system and SDS-PAGE. And the results obtained were as follows; The fractions of retention volume (RV) 0.83ml and RV 1.36ml separated with superose column should stimulate sperm migration and movement. However, RV 0.83 fraction was consisted of complex materials containing RV 1.36 component. RV 1.36 fraction contained a BSA analogue of 67 kilodaltons (Kd) and showed identical peak pattern with BSA fraction V. In conclusion, the protein of 67 Kd in follicular fluid should stimulate sperm migration and movement.

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Improved Fertilization Rate in Human In vitro Fertilization with the Use of a TEST-Yolk Buffer (TEST-Yolk Buffer에 의한 인간 정자의 수정능 증진효과에 관한 연구)

  • Pang, Myung-Geol;Kim, Ki-Chul;Shin, Chang-Jae;Moon, Shin-Yong;Lee, Jin-Yong;Chang, Yoon-Seok
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.1
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    • pp.57-64
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    • 1992
  • The present study was undertaken to clarify the role of TEST-Yolk Buffer(TYB) as a factor for the improvement of human sperm fertility potential. We examined the effects of low temperature capacitation using TYB on sperm motility (%), motility pattern, normal morphology, true acrosome reaction, sperm penetration assay and human in vitro fertilization. Comparing the TYB method and swim-up method, the sperm motility(%) of selected sperm was not significantly different, but statistically significant differences were found in curvilinear velocity, linearity, lateral head displacement, normal morphology(%) and true acrosome reaction(%)(p<0.05). Results obtained from the sperm penetration assay demonstrated that the penetration index and penetration rate were increased significantly(p<0.05) when the spermatozoa were incubated in TYB, as compared with swim-up method. And fertilization of intact human oocytes was more succesful when spermatozoa were pretreated with TYB at $4^{\circ}C$ for 48 hours as compared with swim-up method. Our results show that TYB method have advantages in terms of enhancement of sperm hyperactivation, increased true acrosome reaction, increased ability to penetrate zona-free hamster ova and augmented fertilization of human oocytes, suggesting that TYB is superior in its ability to preserve sperm motility and fertilizing ability.

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Percentage of motile spermatozoa at 22 hours after swim-up procedure: An indicator for intracytoplasmic sperm injection?

  • Inoue, Taketo;Yonezawa, Yukiko;Sugimoto, Hironobu;Uemura, Mikiko;Ono, Yuri;Kishi, Junji;Emi, Nobuyuki;Ono, Yoshiyuki
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.3
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    • pp.157-163
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    • 2016
  • Objective: The decision to use in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI), or split insemination (IVF-ICSI) in the first cycle is based on the number of motile sperm. Hence, total fertilization failure (TFF) often occurs during IVF cycles, despite normozoospermia. To investigate whether the cumulative motile swim-up spermatozoa percentage at 22 hours post-insemination (MSPPI) is an indicator for ICSI, we analyzed TFF, fertilization, blastocyst development, chemical pregnancy, clinical pregnancy, and live birth rates. Methods: This prospective study was performed using data obtained from 260 IVF cycles. At 22 hours after insemination, the remaining swim-up spermatozoa were observed and divided into six groups according to MSPPI (<10%, 10% to <30%, 30% to <50%, 50% to <70%, 70% to <90%, and 90% to 100%). Results: Regardless of the ejaculated motile sperm concentration ($0.6-280{\times}10^6/mL$ motile spermatozoa), the incidence of TFF significantly increased when MSPPI was <10%, and the fertilization rate significantly decreased when MSPPI was <30%. We found that cumulative MSPPI correlated with the cumulative fertilization rate (Spearman correlation, 0.508, p<0.001). Regarding embryo development, we observed no significant differences in the rates of blastocyst development, chemical pregnancy, clinical pregnancy, or live birth among all groups. Conclusion: Our findings suggest that MSPPI is a viable indicator for split IVF-ICSI and ICSI. Taken together, by employing the MSPPI test in advance before IVF, ICSI, or split IVF-ICSI cycles, unnecessary split IVF-ICSI and ICSI may be avoided.

A Study on Education of Life Guard in Jeju-Do (제주도내에서의 수상 인명구조원 교육에 대한 연구)

  • Kang, Soon-Min;Kim, Jae Pil
    • The Korean Journal of Emergency Medical Services
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    • v.5 no.1
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    • pp.73-88
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    • 2001
  • Post these safety tips in any swimming area for all swimmers to read. 1. Always swim with companions. Swim only in area well supervised by lifeguards. 2. Never drink alcohol or use drugs when you're swimming or boating. 3. Always check the water depth. Walk in from shore or ease in from the dock or the edge of the pool. 4. Know the limits of your own swimming abilities. If you're a good swimmer, don't tempt nonswimmers or beginner swimmers to try to keep up with you. 5. Keep any eye on younger swimmers at all times. 6. Learn the proper way to dive in the water safely. Follow guidelines for safe diving. 7. Follow the lifeguards instructions and respect their judgment. Never fake an Emergency. Obey all swimming rules.

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