• Title/Summary/Keyword: Thawing method

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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.

A New Protocol for Effective Cryopreservation of Human Embryonic Stem Cells by a Minimum Volume Cooling Method

  • Kim, Eun-Young;Lee, Keum-Sil;Shin, Hyun-Ah;Park, Sae-Young;Yoon, Ji-Yeon;Kil, Kwang-Soo;Lee, Young-Jae;Kim, Nam-Hyung;Chung, Kil-Saeng
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.98-98
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    • 2003
  • Recently, human embryonic stem (hES) cells have become very important resources for ES cell basic research, cell replacement therapy, and other medical applications; thus, efficient cryopreservation methods for these cells are needed. This study examined whether a newly developed minimum volume cooling (MVC) vitrification method, which was tested through cryopreservation of sensitive bovine oocytes, can be used for freezing hES cells. Feeder-free cultured hES cell (MB03) colonies were mechanically dissected into several small clumps following enzymatic treatment. We compared the freezing efficiency of a slow-cooling method using a cryo-module (0.4-0.6C/min, 20-30 clumps/vial) and MVC vitrification using a modified 0.5-ml French mini-straw designated as a MVC straw (>$20,000{\circ}C$/min, 10 clumps/straw) After thawing, in vitro survival of hES cell clumps was higher for MVC-vitrified cells (80.8%, 97/120) than for slow-cooled cells (38.2%, 39/102). Further, the proliferation rate of surviving MVC-vitrified cells was similar to that of control hES cells from 2 weeks after thawing. In addition, vitrified-thawed hES cells demonstrated a normal karyotype, were positively immunostained for surface marker antibodies (AP, SSEA-4 and TRA-1-60) and the Oct-4 antibody, and could differentiate into all three embryonic germ layer cells in vitro. This result demonstrates that hES cell clumps can be successfully cryopreserved by a newly developed MVC vitrification method without loss of human cell characteristics.

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Establishment of Bovine Ovum Bank : I. Full Term Development of Vitrified Hanwoo (Korean Cattle) In Vitro Matured Oocytes by Minimum Volume Cooling (MVC) Method

  • 김은영;김덕임;이문걸;이종우;이금실;박세영;박은미;윤지연;허영태
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.5-5
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    • 2001
  • This study was to test whether Hanwoo in vitro matured oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5-10 min, exposed in EG30 for 30 sec, each oocytes were individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures [1.0 Msucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS] at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were found to pregnant and three of them were ongoing pregnant by manual palpation at 250 days after transfer. However, among them, two healthy female calves (23 and 25kg) were born. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

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A Study on High-acidity Rubus coreanus Concentrated Vinegar Production Using Freeze Concentration Method (동결농축법을 이용한 고산도 복분자 농축식초 제조에 관한 연구)

  • Sung, Ji-Youn;Lee, Ikheui
    • Journal of the Korea Convergence Society
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    • v.12 no.11
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    • pp.419-426
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    • 2021
  • Until recently, various studies for the production of high-acidity vinegar have been conducted, but there have been few studies on the concentration process of vinegar. In this study, the possibility of introducing freeze-concentration method was investigated for the production of high-quality, high-acidity vinegar. Acidity, pH, specific gravity, and glucose concentration were measured for frozen concentrated vinegar fractions obtained by freezing and thawing three types of Rubus coreanus vinegar with different acidy (7.53, 5.43, and 3.72). Acidity, specific gravity, and glucose concentration were all highest when 15% of the original vinegar was thawed. pH was the lowest when 15% of the original vinegar was melted. As a result of the measurement of cumulative acidity, when 20% of the original vinegar was melted, it was the highest at 9.32, which was 3.89 higher than that of the original vinegar. In this study, it was confirmed that vinegar can be effectively concentrated using the freeze concentration method and at the same time, the thawing ratio can be controlled to effectively obtain vinegar with the desired acidity. However, it is considered that studies such as organic acid and amino acid analysis are needed to determine whether the freeze-concentration method is introduced into the high-acidity concentrated vinegar manufacturing process.

Characteristics of Sand-Silt Mixtures during Freezing-Thawing by using Elastic Waves (탄성파를 이용한 모래-실트 혼합토의 동결-융해 특성)

  • Kang, Mingu;Kim, Sangyeob;Hong, Seungseo;Kim, Youngseok;Lee, Jongsub
    • Journal of the Korean GEO-environmental Society
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    • v.15 no.5
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    • pp.47-56
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    • 2014
  • In winter season, the pore water inside the ground freezes and thaws repetitively due to the cold air temperature. When the freezing-thawing processes are repeated on the ground, the change in soil particle structure occurs and thus the damage of the infrastructure may be following. This study was performed in order to investigate the stiffness change of soils due to the freeze-thaw by using elastic waves. Sand-silt mixtures are prepared with in the silt fraction of 40 %, 60 % and 80 % in weight and in the degree of saturation of 40 %. The specimens are placed into the square freezing-thawing cell by the temping method. For the measurement of the elastic waves, a pair of the bender elements and a pair of piezo disk elements are installed on the cell, and a thermocouple is inserted into soils for the measurement of the temperature. The temperature of the mixtures is decreased from $20^{\circ}C$ to $-10^{\circ}C$ during freezing, is maintained at $-20^{\circ}C$ for 18 hours, is gradually increased up to the room temperature of $20^{\circ}C$ to thaw the specimens. The shear waves, the compressional waves and the temperature are measured during the freeze-thaw process. The experimental result indicates that the shear and the compressional wave velocities after thawing are smaller than those of before freezing. The velocity ratio of after thawing to before freezing of shear wave is smaller than that of the compressional wave. As silt fraction increases from 40 % to 80 %, the shear and compressional wave velocities are gradually increased. This study suggests that the freezing-thawing process in unsaturated soil loosens the soil particle structure, and the shear wave velocity reflects the effect of freezing-thawing more sensitively than the compressional wave velocity.

Changes of the Composition of Nitrogenous Compounds in Globefish Meat Extracts by the Cooking Method (가열조리가 복어 추출물 함질소화합물의 조성에 미치는 영향)

  • Yang, Yeong;Han, Yeong-Sil;Pyeun, Jae-Hyeung
    • Korean journal of food and cookery science
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    • v.6 no.2
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    • pp.85-96
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    • 1990
  • Globefish, Fugu xanthopterus, known to have a severe toxin, is one of the favorite food in Korea and Japan when the toxic part is removed. In this paper, the effect of cooking on the composition of nitrogenous components in the extractives from globefish cooked investigated and the changes of the taste compounds originated from the nitrogenous components in the extractives were discussed. When the sample fish was analysed for general composition, drip amount and pH by the different methods of thawing, the method effective method was the running water thawing. Total nitrogen content in raw globefish and the frozen globefish was not different, and the nitrogen content was increased with the heat treatment. It seemed that the nitrogen content was higher in the extract from the boiled globefish than that of the steamed globefish. Taurine, lysine, glycine and alanine were occupied about 70% of the total free amino acids. Total free amino acid content was higher in the extracts from the frozen sample than those from the raw sample. The amount of free amino acids was increased when the globefish soup cooked under the direct-heat cooking than in the microwave oven-heat cooking. Among nucleotides in the extracts from the thawed and cooked fishes, IMP and inosine contents were increased, and the both components were decreased with the heating time and by the heating method. Tne content of total creatinine-nitrogen were 50% of the total nitrogen content of the extracts, but the concentration of glycinebetaine, TMA and TMAO were only a few amounts. It could be concluded that total creatinine components, including free amino acids such as taurine, lysine, glycine and alanine, and IMP might be the important components contributing to the taste of the cooked globefish.

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Evaluation of Horizontal Force on Pile Shaft Surrounded by Vertical PET Aggregate Layer for Fluid Machinery Structure Installation in Cold Region's Plant (동토 플랜트 유체기계 구조물 설치를 위한 PET 골재적용 말뚝의 주면작용 수평력 평가)

  • Ji, Subin;Jang, Sung Min;Hwang, Soon Gap;Lee, Kicheol;Kim, Dongwook
    • The KSFM Journal of Fluid Machinery
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    • v.19 no.3
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    • pp.43-47
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    • 2016
  • Pile foundations constructed on extremely cold regions cause serviceability problems of superstructures from repeated actions of ground freezing and thawing. Oil sand module plants are mainly constructed on seasonal frozen ground. Due to the freezing and thawing actions of grounds, vertical movements of piles have been observed. To solve these erratic pile movement problems, thin vertical layer of PET aggregates is installed around the pile shaft to prevent potential unfavorable pile movements. There is no known method to calculate "thin PET aggregate layer" -surrounded pile shaft resistance (capacity) against vertical loads; therefore, this experimental research is conducted. Specifically, in this study, horizontal (normal) pressures on pile shaft were assessed varying PET aggregate layer thickness based on the experiment.

Evaluation for Moisture Susceptibility of Asphalt Mixtures using Non-Destructive Impact Wave (비파괴 충격파를 이용한 아스팔트 공시체의 수분민감도 평가)

  • Jang, Byung Kwan;Kim, Do Wan;Mun, Sung Ho;Jang, Yeong Sun
    • International Journal of Highway Engineering
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    • v.15 no.3
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    • pp.53-63
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    • 2013
  • PURPOSES : This study is to evaluate moisture susceptibility of asphalt mixtures by using non-destructive impact wave and to determine durability so as to decrease the gap between before and after freezing in the future. METHODS : Using non-destructive impact wave, this study is to determine the dynamic modulus of asphalt specimen. Furthermore, the results obtained from two experiment accelerometers are used for the dynamic modulus determination. The dynamic moduli of specimens are compared with those of the freezing-thawing specimens. RESULTS : Test results showed that the dynamic modulus before freezing and thawing environment loads at each temperature dropped about 3.7% after the environmental loads. Furthermore, correlation analysis indicates that transition of dynamic modulus at each point is about 89.59%. CONCLUSIONS: Evaluation of asphalt mixtures using non-destructive impact wave has excellent repeatability and simple equipment for the test. Consequently, the method in the study will be useful for evaluating the characteristics of a various asphalt mixtures.

Development of Cryopreservation System using Shoot-Apex in Yam (Dioscorea batatas)

  • Shin Jong-Hee;Kang Dong-Kyoon;Bae Jeong-Suk;Lee Bong-Ho;Sohn Jae-Keun
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.43-50
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    • 2006
  • The goal of this research was to develop an efficient cryopreservation protocol for germplasms of yam (Diosorea batatas), that were cultivated in Korea. Comparative studies with four other cryogenic techniques and subsequent experiments for shoot regrowth were conducted. in vitro-grown shoot-apices of the D. batatas were successfully cryopreserved by encapsulation-dehydration. The maximum survival of shoot-apices could be achieved when the precultured (with 0.3 M of sucrose for one day) and encapsulated (with a 3%(w/v) Na-alginate solution) apices were dehydrated for $3.5{\sim}4\;h$ prior to direct immersion in LN (liquid nitrogen). The frequency of regrowth rate of cryopreserved apices was not decreased during 3-month storage period. The thawing method markedly affected survival of the cryopreserved apices, and thawing at $40^{\circ}C$ for 3 min produced the best results. When cryopreserved apices were post-cultured on the post-culture medium (MS), supplemented with $0.2mgl^{-1}$ of BA ($N_6$-benzyladenine) and $0.2mgl^{-1}$ of kinetin, they showed direct shooting without callusing.

Studies on the Rapid Freezing of Mouse Embryo II. Effects of the Development Stage and Seeding on the Mouse Embryo Survival of Rapid Freezing (Mouse 수정란의 급속동결에 관한 연구 제II보 Mouse수정란 급속동결에 있어서 수정란의 발육단계와 식빙(seeding)이 생존율에 미치는 영향)

  • 강만종;김영훈;문성호;김중규
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.141-148
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    • 1989
  • The effects of seeding method and optimum time for freezing embryos according to the developmental stages on embryo survival rates after rapid freezing were determined using the FDA-test. The summarized results are as follows : 1. In the rapid freezing of embryos, the sucrose added medium together with Co-seeding or non-seeding showed the FDA scores of 4.67 and 4.20, respectively, but, raffinose addition obtained FDA scores of 4.27 and 3.97. 2. The developmental stage of embryos at freezing was most critical on the survival of embryos after thawing. Higher FDA scores were obtained in the order of blastocyst stage(4.94), morula stage(3.82) and ealy stage(2.65) in sucrose added medium. The same trend was observed in the raffinose added medium with an order or 4.91, 4.47 and 2.32. 3. Microscopic study of embryo before freezing and post-thawing indicated that the embryo showed shrinkage within 5 minutes after the embryo was transfer to the freezing medium. When thawed embryo was tranfered to the dilution medium, swelling of the embryo was observed and there after it reshrank indicating the removal of cryoprotectant from the embryo. The size of the embryo recovered to the original state when it was moved into a PBS-solution.

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