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Systems for Production of Calves from Hanwoo(Korean Cattle) IVM/IVF/IVC Blastocyst III. Vitrification and One-Step Dilution of Hanwoo Blastocyst (체외생산된 한우 배반포기배로부터 송아지 생스을 위한 체계 III. 한우 배반포기배의 초자화 동결과 1단계 융해)

  • Kim, E.Y.;Kim, D.I.;Park, N.H.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.293-301
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    • 1999
  • This study was to examine whether Hanwoo IVM/IVF/IVC blastocyst can be successfully survived in vitro/in vivo after vitrification and one-step dilution. For vitrification, blastocysts were serially exposed in glycerol (G) and ethylene glycol(EG) mixtures[10% (v Iv) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.] which is diluted in 10% FBS added D-PBS. And then they were loaded in the straw, placed in cold nitrogen vapor for 3 min. and plunged into L$N_2$(-196$^{\circ}C$). One-step dilution within the straw was done in $25^{\circ}C$ and 36$^{\circ}C$ water for about 5 min. and 3 min., respectively. Recovered embryos after one-step dilution were cultured in cumulus cell mono-layered drop for 48 h or were transferred into recipient cows. When the embryo survival in vitro was assessed to re-expanded and hatched rates at 24 hand 48 h after one-step dilution, the results of vitrified group (85.4, 43.8%) was high, although these results were significantly lower than normal development (100.0, 63.3%) of control group, respectively (P<0.001, P<0.05). When in vitro survival of vitrified groups according to developmental stage was compared, the results of fast developed embryos (expanded blastocyst and early hatching blastocyst stage) were significantly higher than those of delayed developed one (early blastocyst stage) after one-step dilution (early hatching: 88.0, 48.0%: expanded: 81.1, 45.3%; early: 66.7, 14.3%) (P<0.05). Also, in case of in vitro survival of vitrified groups according to embryo age (day 7, 8 and 9), when embryo age was younger, in vitro survival was significantly higher (day 7: 67.3, 34.5%; day 8: 76.9, 40.7%; day 9: 60.9, 23.9%)(P<0.05). Finally, when in vivo development potential of vitrified and one-step diluted Hanwoo blastocysts was examined, 4 of 8 recipient (50%) cows became confirmed pregnant. These results demonstrated that our vitrification and one-step dilution technique can be applied easily and effectively on field trial without the equipment and embryological skills required for conservative dilution and transfer.

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Effect of Season on Testis Function and Freezing and Fertilizing Ability of Spermatozoa in Korean Native Goat I. Seasonal Changes in Semen Characteristics and Freezing and Penetrating Ability of Sperm (한국재래산양에서 계절이 정소기능, 정자의 내동성 및 수정능력에 미치는 영향 I. 정액성상과 정자의 내동성 및 난자침입능력의 계절적 변화)

  • Kim, C.K.;Chung, Y.C.;Kim, K.S.;Yoon, J.T.;Lee, J.H.;Chung, Y.H.;Choi, S.H.;Kim, H.Y.;Kim, S.;Kwon, C.J.
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.311-323
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    • 1994
  • This study was conducted to observe seasonal and individual changes in semen characteristics and sperm freezability, and sperm penetration into zona-free hamster eggs in Korean native goats. Buck response and change in semen characteristics to electrical stimulations was evaluated for four seasons throughout 2 years and percentage of motile sperm and normal apical ridge acrosome was investigated after equilibration and thawing for 4 seasons with 5 bucks. Sperm penetration rate was evaluated for 4 bucks. 1. Probe insertion at depth of 7cm and repeated stimulation for 3 sec was more effective(P<0.05) in buck response and semen collection than those of other conditions. 2. Semen characteristics from electrojaculation was signficantly(P<0.005) higher in spring and fall for semen volume, in spring and summer for sperm concentration and in fall for sperm motility than those in other seasons, respectively. However, there were no differences in total sperm among seasons. 3. Buck response to electrical stimulation showed significant difference(P<0.05) among individuals in all 3 seasons except winter. Significant individual difference in semen volume was only in spring and summer, but there was no indivudual difference in sperm concentration and total sperm in all season. 4. Washing of semen before freezing treatment was greatly(P<0.05) beneficial to sperm motility after thawing, no matter whether ejaculates exhibit egg yolk coagulation or not. 5. Sperm motility after glycerol equilibration was significantly(P<0.05) low in summer semen and motility after thawing was greatly(P<0.05) higher in winter semen than in other seasons. Freezability of unwashed sperm was significantly difference among bucks, but a yearly freezability of washed sperm after chilling and thawing were no differences among bucks and percentage of normal apical ridge acrosome were not different among seasons and bucks. 6. There was no significant difference in sperm motility after thawing between egg yolk levels in summer, although 20% level gave more higher motility than 5% level. 7. In summer, 3.2% glycerol and 3-h equilibration gave greatest percentage(P<0.05) of sperm motility and normal apical ridge acrosome after thawing. 8. Sperm penetration rate into zona-free hamster eggs was not different between bucks and seasons. Overall, it is concluded that to obtain maximum sperm output and successive semen freezing by electrojaculation method, buck selection with good response in all season could be basically considered and that seasonal effect on sperm freezability was more greater than that of individual bucks.

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Systems for Production of Calves from Hanwoo(Korean Native Cattle) IVM/IVF/IVC Blastocyst I. Hanwoo IVM/IVF /IVC Blastocyst Cryopreserved by Vitrification (체외생산된 한우 배반포기배로부터 송아지 생산을 위한 체계 I. 체외생산된 한우 배반포기배의 초자화 동결보존)

  • Park, S. P.;Kim, E. Y.;Kim, D. I.;Park, N. H.;Y. S. Won;S. H. Yoon;K. S. Chung;J. H. Lim
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.349-357
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    • 1998
  • This study was to investigate whether the viability of Hanwoo IVM/IVF/IVC blastocysts was maintained after vitrification and thawing. In vitro produced Hanwoo blastocysts were vitrified by two-step method: equilibrated in EG20 for 3 min, and then exposed in EFS40 [40% ethylene glycol (EG), 18% ficoll and 10.26% sucrose in mDPBS containing 10% FBS ]and vitrified in L$N_2$for 30 - 45 sec. After thawing, in vitro survival was assessed as the re-expanded and hatched rates at 24 hand 48 h, respectively. The results obtained in these experiments were summarized as follows: From the 12 replicates, 52.5% of Hanwoo blastocysts were produced in vitro at day 7 after IVF. When the effects of freezing solution to the embryo survival were examined, there is no significant toxicity in exposure (100.0, 73.8%) compared to that af control group (100.0, 87.0%). However, when embryos were vitrified, high survival (86.2, 55.4%) was obtained although it was significantly lower than those of exposure and control group (p<0.05). When the in vitro survival of vitrified embryos according to developmental stage and culture day were examined, it showed that more advanced embryo stage exhibited a significantly higher survival rate irrespective of culture day (p<0.05). Also, even in the same development stage, the in vitro survival of day 7 embryos (re-expanded: 75.0~87.5%, hatched: 21.4~66.7%) was higher than those of day 8 embryos(re-expanded: 58.6~78.3%, hatched: 10.3~52.2%). Therefore, these results suggested that in vitro produced Hanwoo blastocysts can be successfully cryopreserved by simple two-step vitrification method using EFS40 freezing solution, particularly at the expanded and early hatching blastocyst stage regardless of embryo culture duration (day 7 or day 8 after IVF).

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Development of Respiration Gating RT Technique using Moving Phantom and Ultrasound Sensor: a feasibility study (동 팬텀과 초음파 센서를 이용한 호흡운동 조절 방사선치료 기술 개발)

  • Lee, Suk;Lee, Sang-Hoon;Shin, Dong-Ho;Yang, Dae-Sik;Choi, Myung-Sun;Kim, Chul-Yong
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2004.11a
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    • pp.122-125
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    • 2004
  • In radiotherapy of tumors in liver, enough planning target volume (PTV) margins are necessary to compensate breathing-related movement of tumor volumes. To overcome the problems, this study aims to obtain patients' body movements by using a moving phantom and an ultrasonic sensor, and to develop respiration gating techniques that can adjust patients' beds by using reversed values of the data obtained. The phantom made to measure patients' body movements is composed of a microprocessor (BS II, 20 MHz, 8K Byte), a sensor (Ultra-Sonic, range 3 cm ${\sim}$3 m), host computer (RS232C) and stepping motor (torque 2.3Kg) etc., and the program to control and operate it was developed. The program allows the phantom to move within the maximum range of 2 cm, its movements and corrections to take place in order, and x, y and z to move successively. After the moving phantom was adjusted by entering random movement data(three dimensional data form with distance of 2cm), and the phantom movements were acquired using the ultra sonic sensor, the two data were compared and analyzed. And then, after the movements by respiration were acquired by using guinea pigs, the real-time respiration gating techniques were drawn by operating the phantom with the reversed values of the data. The result of analyzing the acquisition-correction delay time for the three types of data values and about each value separately shows that the data values coincided with one another within 1% and that the acquisition-correction delay time was obtained real-time (2.34 ${\times}$ 10$^{-4}$sec). This study successfully confirms the clinic application possibility of respiration gating techniques by using a moving phantom and an ultra sonic sensor. With ongoing development of additional analysis system, which can be used in real-time set-up reproducibility analysis, it may be beneficially used in radiotherapy of moving tumors.

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Development of Respiration Gating RT Technique using Moving Phantom and Ultrasound Sensor: a feasibility study (동 팬텀과 초음파 센서를 이용한 호흡운동 조절 방사선치료 기술 개발)

  • Lee Suk;Lee Sang Hoon;Shin Dongho;Yang Dae Sik;Choi Myung Sun;Kim Chul Yong
    • Radiation Oncology Journal
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    • v.22 no.4
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    • pp.316-324
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    • 2004
  • Purpose : In radiotherapy of tumors in liver, enough planning target volume (PTV) margins are necessary to compensate breathing-related movement of tumor volumes. To overcome the problems, this study aims to obtain patients' body movements by using a moving phantom and an ultrasonic sensor, and to develop respiration sating techniques that can adjust patients' beds by using reversed values of the data obtained. Materials and Methods : The phantom made to measure patients' body movements is composed of a microprocessor (BS II, 20 MHz, 8K Byte), a sensor (Ultra-Sonic, range $3\~3$ m), host computer (RS232C) and stepping motor (torque 2.3 Kg) etc., and the program to control and operate it was developed. The program allows the phantom to move within the maximum range of 2 cm, its movements and corrections to take place In order, and x, y and z to move successively. After the moving phantom was adjusted by entering random movement data (three dimensional data form with distance of 2 cm), and the phantom movements were acquired using the ultra sonic sensor, the two data were compared and analyzed. And then, after the movements by respiration were acquired by using guinea pigs, the real-time respiration gating techniques were drawn by operating the phantom with the reversed values of the data. Results : The result of analyzing the acquisition-correction delay time the three types of data values and about each value separately shows that the data values coincided with one another within $1\%$ and that the acquisition-correction delay time was obtained real-time $(2.34{\times}10^{-4}sec)$. Conclusion : This study successfully confirms the clinic application possibility of respiration gating techniques by using a moving phantom and an ultrasonic sensor. With ongoing development of additional analysis system, which can be used in real-time set-up reproducibility analysis, it may be beneficially used in radiotherapy of moving tumors.

A Kinematics Analysis of Uchi-mata(inner thigh reaping throw) by Kumi-kata types and Two different Opponent's Height in Judo[II] (유도 맞잡기 타입과 받기의 신장에 따른 허벅다리걸기의 Kinematic 분석[2])

  • Kwon, Moon-Seok;Kim, Eui-Hwan;Cho, Dong-Hee
    • Korean Journal of Applied Biomechanics
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    • v.12 no.2
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    • pp.143-157
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    • 2002
  • This study was to analyze the kinematic variables when the subjects performing Uchi-mata(inner thigh reaping throw) by Kumi-kata types((How to grasp A or B?) and two different opponent's height in Judo. Kinematic variables were temporal, posture. Data analysis was collective comparison of two-way ANOVA, t-test by type A&B and two different opponent's height. There were significant difference of Kumi-kata types(p<.05) in the time elapsed on Kake phase(KP : throwing phase) and hip, knee, ankle-angle of the attacking foot in the 1st stage of KP and knee, ankle-angle of the attacking foot and hip, knee ankle-angle of the supporting foot in the 2nd stage of KP. There were significant difference of two opponent's(p<.05) in the time elapsed on KP and hip-angle of the supporting foot in 1st stage of KP. Therefore, the interaction effect(p<.05) were in the time elapsed on KP and hip-angle of the supporting foot in the 2nd stage of KP. So, It could be suggested that Judoka hold on the part-behind neck lapel(type A) at the sleeve with the other of Judogi jacked when opponent's height was short. Because the time elapsed on KP of type B was not so fast as type A(p<.05) during performed Uchi-mata, and also the bigger hip-angle of the supporting foot in the 2nd stage of KP grew, the faster the time elapsed on KP became.

A Case Study on Center of Gravity Analysis when Performing Uchimata by Posture and Voluntary Resistance Levels of Uke in Judo[ll] (유도 허벅다리걸기 기술발휘 시 받기의 자세와 저항수준에 따른 중심변인 분석 사례연구[II])

  • Kim, Eui-Hwan;Kim, Sung-Sup;Chung, Chae-Wook
    • Korean Journal of Applied Biomechanics
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    • v.15 no.1
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    • pp.237-257
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    • 2005
  • It was to study as a following-research of "A Case Study on Center of Gravity(COG) Analysis when Performing Uchimata(inner thigh reaping throw) by Posture and Voluntary Resistance Levels(VRL) of Uke in Judo[I]". The purpose of this study was to analyze the COG variables when performing uchimata(inner thigh reaping throw) by two postures and voluntary resistance levels(VRL) of uke(reciver) in Judo. The subjects, who were one male judoka(YH) for 1992 Barcelona Olympic Games Olympian(silver medalist), and one male trainee; Y.I.University representative member (SDK), and were filmed on two S-VHS 16mm video cameras(60fields/sec.) through 3-dimensional motion analysis methods, that postures of uke were shizenhontai (straight natural posture) and jigohontai(straight defensive posture), VRL of uke were 0% and 100%, respectively. The kinematical variable was COG variable, distance of COG, and distance of resultant COG between uke and tori(the thrower), velocity and acceleration of COG. The data of this study collection were digitized by SIMI Motion Program computed the mean values and the standard deviation calculated for each variables. When performing uchinmata according to each posture and VRL of uke and classifying. From the data analysis and discussion, the conclusions were as follows : 1. Displacement of COG Subject YH, COG was the highest in kuzushi(balance -breaking), vertical COG was low when following in tsukuri(positioning; set-up), kake(application; execution), and COG was pattern of same character each postures and resistance, respectively. Subject SDK, COG was low from kumikata(engagement positioning) to kake, and COG was that each postures and resistance were same patterns, respectively. Subject YH, SDK, each individual, postures and resistance, vertical COG was the lowest in kake phase, when performing. 2. Distance of COG between uke and tori The distance of COG between uke and tori when performing, subject YH was $0.64{\sim}0.70cm$ in kumikata, $0.19{\sim}0.28cm$ in kake, and SDK was $0.68{\sim}0.72cm$ in kumikata, $0.30{\sim}0.42\;cm$ in kake. SDK was wider than YH. 3. Distance of resultant COG between uke and tori The distance of resultant COG between uke and tori when performing, subject YH was $0.27{\sim}0.73cm$ from kumikata to kake. and SDK was $0.14{\sim}0.34cm$ in kumikata, $0.28{\sim}0.65cm$ in kake. Jigohontai(YH:$0.43{\sim}0.73cm$,SDK:$0.59{\sim}0.65cm$) was more moved than shizenhontai(YH:$0.27{\sim}0.53cm$, SDK: $0.28{\sim}\;0.34cm$). 4. Velocity of COG The velocity of COG when performing uchimata, subject YH was fast anterior-posterior direction in kuzushi, ant.-post. and vertical direction fast in tsukuri and kake. SDK was lateral, ant.-post. and vertical direction in kuzushi, ant.-post. and vertical direction in tsukuri and ant.-post. direction in take, respectively. 5. Acceleration of COG The acceleration of COG when performing uchimata, The trend of subject YH was showed fast vertical direction in kuzushi and tsukuri, ant.-post. and vertical direction fast in kake. The trends of SDK showed lateral direction in kuzushi, lateral and ant.-post. direction in tsukuri and ant.-post. direction in kake, respectively.

Implementation of a Self Controlled Mobile Robot with Intelligence to Recognize Obstacles (장애물 인식 지능을 갖춘 자율 이동로봇의 구현)

  • 류한성;최중경
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.40 no.5
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    • pp.312-321
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    • 2003
  • In this paper, we implement robot which are ability to recognize obstacles and moving automatically to destination. we present two results in this paper; hardware implementation of image processing board and software implementation of visual feedback algorithm for a self-controlled robot. In the first part, the mobile robot depends on commands from a control board which is doing image processing part. We have studied the self controlled mobile robot system equipped with a CCD camera for a long time. This robot system consists of a image processing board implemented with DSPs, a stepping motor, a CCD camera. We will propose an algorithm in which commands are delivered for the robot to move in the planned path. The distance that the robot is supposed to move is calculated on the basis of the absolute coordinate and the coordinate of the target spot. And the image signal acquired by the CCD camera mounted on the robot is captured at every sampling time in order for the robot to automatically avoid the obstacle and finally to reach the destination. The image processing board consists of DSP (TMS320VC33), ADV611, SAA7111, ADV7l76A, CPLD(EPM7256ATC144), and SRAM memories. In the second part, the visual feedback control has two types of vision algorithms: obstacle avoidance and path planning. The first algorithm is cell, part of the image divided by blob analysis. We will do image preprocessing to improve the input image. This image preprocessing consists of filtering, edge detection, NOR converting, and threshold-ing. This major image processing includes labeling, segmentation, and pixel density calculation. In the second algorithm, after an image frame went through preprocessing (edge detection, converting, thresholding), the histogram is measured vertically (the y-axis direction). Then, the binary histogram of the image shows waveforms with only black and white variations. Here we use the fact that since obstacles appear as sectional diagrams as if they were walls, there is no variation in the histogram. The intensities of the line histogram are measured as vertically at intervals of 20 pixels. So, we can find uniform and nonuniform regions of the waveforms and define the period of uniform waveforms as an obstacle region. We can see that the algorithm is very useful for the robot to move avoiding obstacles.

Sister Chromatid Exchange (SCE) Frequency and In Vitro Development of Mouse Zygote Cryopreserved by Vitrification (초자화 동결에 의한 생쥐 1-세포기배의 체외 발달과 SCE 빈도)

  • Kim, M.K.;Baik, C.S.;Uhm, S.J.;Kim, E.Y.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.379-384
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    • 1996
  • This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.

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Cryopreservation of Bovine IVM/IVF/IVC Blastocysts by Vitrification (체외성숙, 체외수정 및 체외배양에서 생산된 소 배반포기배의 초자화 동결)

  • Nam, H.K.;Kim, E.Y.;Lee, K.S.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.231-238
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    • 1999
  • The objective of this study was to examine the effect of developmental stage and embryo age of in vitro produced bovine blastocysts after vitrification and thawing. In vitro cultured day 8 blastocysts after IVF were equilibrated 20% ethylene glycol (EG) for 3 min. and were vitrified using EFS40, which is consisted of 40% EG, 18% ficoll, 0.3M sucrose and 10% FBS added in mDPBS for 30 sec. before being plunged into $LN_2$. Also, survival in vitro was assessed by re-expansion and hatching or hatched at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) When the embryos were cultured for 8 day after IVF, 41.0% of the cleaved embryos developed to the blastocysts (early; 7.6%, expanded; 22.9%, hatching; 4.6% and hatched; 5.9%). 2) When the embryos were exposed or vitrified to the freezing solution, the re-expansion of vitrified embryos (73.3%) was significantly lower than that of control and exposed embryos (100, 97.0%) (p<0.05). But the formation rate of hatching or hatched blastocysts of vitrified embryos (66.7, 46.7%) at 48h after thawing was similar to that of exposed embryos (66.7, 39.4%) but not control (100, 100%) (p<0.01). However, in the total cell numbers of those developed hatched blastocysts, there were not significantly different among the treatment groups. 3) When the embryo survival rates by different developmental stage were examined, the re-expansion was not different among the groups $(64.5{\sim}75.6%)$. After warming 48 h, the hatching and hatched formation of early blastocysts (25.8, 9.7%) was significantly lower than those of expanded (69.7, 39.4%) and hatching blastocysts (53.3, 43.3%) (p<0.05). 4) In addition, when the expanded blastocysts at day 7, 8 and 9 were vitrified, the re-expansion of day 8 and 9 embryos was significantly lower than that of day 7 (day 7; 93.9%, day 8; 75.8% and day 9; 87.5%) (p<0.05). However, the rates of development to hatched blastocysts were no difference among the groups (day 7; 36.4%, day 8; 36.4% and day 9; 31.3%). These results suggested that in vitro produced expanded or hatching blastocysts can be efficiently cryopreserved by the two-step vitrification method using EFS40.

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