• 제목/요약/키워드: thawing methods

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Efficacy of Frozen-Thawed ET in Patients with Old Age or Non-Pregnant in Fresh ET Cycles (고령 환자와 신선주기 배아이식에서 임신에 실패한 환자에서 동결-융해 배아이식의 효용성)

  • Choi, Su Jin;Lee, Sun Hee;Song, In Ok;Koong, Mi Kyoung;Kang, Inn Soo;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.33 no.4
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    • pp.237-243
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    • 2006
  • Objective: The aim of this study was to evaluate the efficacy of frozen-thawed ET in poor prognosis patients such as the old age (38~44 years; OA group) and the patients who did not achieve clinical pregnancy with the first fresh ET cycle (non-pregnant patients; NP group). Methods: Laboratory and clinical data were collected from fresh and frozen-thawed ET cycles of OA and NP group. Controlled ovarian hyperstimulation (COH) and conventional insemination or ICSI, in vitro culture and ET were performed by routine procedures. Supernumerary embryos were frozen by the slow freezing method, and frozen embryos were thawed by the rapid thawing method. Embryo development, pregnancy and implantation rates were statistically analyzed by Student t-test and chi square test Results: Mean ages were similar between fresh ET ($40.0{\pm}1.8$ years, n=206) and frozen-thawed ET ($39.9{\pm}1.9$ years, n=69) cycles in OA group. However, the clinical pregnancy and implantation rate of subsequent frozen-thawed ET significantly higher than those of fresh ET cycles (29.0% and 11.2% vs. 16.5% and 7.0%, p<0.05). In NP group, there was no difference in the mean age between fresh ET ($31.2{\pm}2.3$ years, n=40) and frozen-thawed ET ($31.9{\pm}3.1$ years, n=119) in subsequent cycles. The clinical pregnancy and implantation rates were similar between the subsequent fresh ET (42.5% and 22.6%) and the frozen-thawed ET (40.3% and 18.8%). Conclusion: In old age patients, higher pregnancy rate of frozen-thawed ET compared to fresh ET cycles in this study. It may be related that better uterine environments for implantation in frozen-thawed ET cycles than that of non-physiological hormonal condition in uterus of fresh COH cycles.

Effects of the Artificial Shrinkage and Assisted Hatching Before Vitrification on the Development of the Vitrified Mouse Expanding Blastocysts (유리화동결 전 인공수축과 보조부화술이 융해 후 생쥐 포배아의 발달에 미치는 영향)

  • Jo, Deok-Hyeon;Ko, Gyoung-Rae;Jung, Ji-Hye;Choi, Jong-Ryeol;Joo, Jong-Kil;Lee, Kyu-Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.275-283
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    • 2008
  • Objective: This study was conducted to investigate the effects of the artificial shrinkage and assisted hatching (PZD; patial zona dissetion) before vitrification on the development of vitrified mouse expanding blastocyst. Methods: Mouse 2-cell embryos were collected and cultured in G1.1 and G2.2 to expanding blastocyst. For artificial shrinkage (AS) the micro injection pipette was inserted into blastocoele cavity and blastocoele fluid was aspirated. For assisted hatching (AH) PZD method was used. Control group was -AS/-AH and treatment groups were -AS/+AH, +AS/-AH and +AS/+AH. After AS and AH mouse blastocysts were equillibrated in G10 and G10E20 for 3 mins, respectively, and vitrified in G25E25 after loading on capped pulled-straw. Vitrified mouse blastocysts were thawed and cultured for 24 hrs. The survival and hatching rate was compared among one control and three treatment groups. Results: The survival rates were 99%, 92% in +AS/+AH and +AS/-AH groups and 54%, 58% in -AS/-AH and -AS/+AH group, respectively. The survival rate was significantly higher in +AS group than in -AS group (p<0.01). Hatching rates were 34%, 96% in -AS/-AH and -AS/+AH groups and 41%, 100% in +AS/-AH and +AS/+AH, respectively. The hatching rates was higher in +AH group than in -AH group (p<0.01). After thawing recovery rates were 100%. Loading on capped pulled-straw, that is effective and useful method on vitrification. Conclusion: This study showed that artificial shrinkage of blastocoele cavity and assisted hatching (PZD) significantly improved the development of the vitrified mouse expanding blastocysts.

A study on short-term stability of recombinant protein A (Recombinant protein A의 short-term stability에 관한 연구)

  • Kim, Yoo-Gon;Lee, Woo-Jong;Won, Chan-Hee;Kim, Yong-Hee;Yun, Ji-Sun;Hong, Min-Seon;Shin, Chul-Soo
    • Analytical Science and Technology
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    • v.24 no.3
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    • pp.193-199
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    • 2011
  • The purpose of this experiment is to evaluate the stability of products according to the storage methods, the period of use, and the diurnal variations through the short term stability experiment of recombinunt protein A (rProtein A) produced in AP Tech Co. That is, we investigated how long the stability of the products would last, when we used the samples frozen at $-20^{\circ}C$, which is one of the storage conditions of the produced rProtein A and then kept them refrigerated at $4^{\circ}C$. The experiment was conducted for 8 weeks and 6 experiment points were established. The experiment was done by thawing the samples frozen at $-20^{\circ}C$ at room temperature, and then refrigerating them at $4^{\circ}C$. In addition, experiments for endotoxin, bioburden, HPLC purity, and concentration were conducted. As a result of the experiment, 0.5 EU/mg endotoxin was detected both at the beginning and at the 8th week and bioburden was not analyzed. In the case of purity, it showed 99.23~99.90% at 210 nm (RSD% 0.23%) and 100% at 280 nm, which meant the change into other materials didn't happen and there was no material degradation characteristics. Finally, we also found the fact that the concentration stayed stable at 55.15 mg/mL (RSD% 0.55%) both at the beginning and at the end. From the experiment results, we were able to conclude that the stability at the condition to store rProtein A at 4 oC for 8 weeks was procured without producing microorganisms or having material degradation characteristics.

Effect of Cryopreservation Day on Pregnancy Outcomes in Frozen-thawed Blastocyst Transfer (동결 해동한 포배 이식에 있어서 동결시기가 임신결과에 미치는 영향)

  • Kim, Hyun-Jung;Kim, Chung-Hyon;Lee, Joong-Yeup;Kwon, Jae-Hee;Hwang, Do-Yeong;Kim, Ki-Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.57-64
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    • 2010
  • Objectives: Likewise fresh cycle, it is also important to select right blastocysts for transfer in purpose of improving the pregnancy and implantation rates in frozen-thawed embryo transfer (ET) cycles. To investigate the relationship between the developmental velocity at the time of cryopreservation and pregnancy rates, we compared pregnancy rates between the day 5 cryopreservation group and the day 6 cryopreservation group. Methods: Transfers of frozen-thawed blastocysts which had been cryopreserved by vitrification on day 5 or day 6 were performed between January 2006 and June 2007. Ethylene glycol, DMSO, and pull and cut straws were used for vitrification and artificial shrinkage was done in expanded blastocysts. Thawing was performed on the day before transfer and thawed blastocysts were cultured in for 15~18 hrs in Quinn's blastocyct media. Blastocyst survival was assessed before transfer and post-thaw survival was defined as >50% of cells remaining intact and blastocoele re-expansion by the time of transfer. Results: Transfers of thawed blastocyst had been cryopreserved on day 5 were 52 cycles and 41 transfer cycles were cryopreserved on day 6. Patient characteristics, the number of transferred embryos and the survival rate of thawed blastocysts were not different in each cryopreservation day. But the biochemical pregnancy, clinical pregnancy, ongoing pregnancy, and implantation rate were significantly high in transfer of frozen-thawed blastocyst which were cryopreserved on day 5. Conclusions: The clinical pregnancy and implantation rate of day-5 blastocyst showed significantly higher than those of day-6 blastocyst in frozen-ET cycles. This result indicated that developmental rate of blastocyst at cryopreservation time in frozen-thawed cycle is discriminative marker of pregnancy outcome as like in fresh cycle.

Factors Required to Sustain Pastoral Farming Systems and Forage Supply In Winter-Cold Zones in Canada (캐나다의 동계한냉지역에 있어서 초지개발과 조사료 공급의 활성화에 필요한 요인)

  • Kunelius, H.T.;Fraser, Joanna
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.12 no.3
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    • pp.3-12
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    • 1992
  • Forage grasses and legumes ar$\varepsilon$ the mam component of livestock diets in Canada. There are over 30 million ha of grassland in Canada and there is a large, undeveloped land base in fringe areas suitable for forage production. The short growing s season limits the grassland farming to the southern p parts of Canada. The win!er season is long and in most parts of Canada cold temperatures, fr$\varepsilon$ezmg, and thawing, and diseases exert sever$\varepsilon$ stress on overwintering forage plants. The development of persistent cultivars is essential for sustained production particularly in the fringe areas with short growmg s$\varepsilon$ason. The seasonality of dry matter production is a result of high growth rates in early summ$\varepsilon$r and low dry matter accumulation in late summer and fall. Innovative management practIces a and cultivars with improved regrowth capacity are n necessary to overcome such skewed production pattern and to extend effiectlVe grazmg season l Improved pasture production is an important part of reducing costs in livestock operations and remaining competitive. It is suggested that applying available technology would increase pasture productivity and reduce d$\varepsilon$pendence on stored feeds thus improving profitability of small producers in particeular. Reducing nutrient losses during harv$\varepsilon$stmg, s storage, and feeding is essential for improved production efficiency during confinement. The devclopment of low cost and labor saving methods of ensiling is critical for improved efficiency and profitability of forage based enterprises Livestock industries must respond to consumer preferences for low fat and cholesterol foods. Research and development of entire production systems is emphasized for dev$\varepsilon$loping viabl$\varepsilon$ enterprises. It is increasingly difficult to secure resources for r$\varepsilon$search, education, and extension, and alliane$\varepsilon$s and cooperation must expand among organizations with interests in forage based livestock systems.

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EVALUATION OF THE VIABILITY OF PERIODONTAL LIGAMENT CELL IN RAT TEETH USING SLOW CRYOPRESERVATION METHOD WITH MAGNETIC FIELD (자기장 저속 냉동보관법을 이용한 쥐 치아 치주인대세포의 활성도 검사)

  • Ahn, Hyun-Jung;Kim, Eui-Seong;Kim, Jin;Kim, Duck-Won;Kim, Ki-Yeol;Lee, Chan-Young;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.33 no.4
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    • pp.332-340
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    • 2008
  • The purpose of this study was to evaluate the viability of periodontal ligament cell in rat teeth using slow cryopreservation method with magnetic field through MTT assay and TUNEL test. For each group, 12 teeth of 4 weeks old white female Sprague-Dawley rat were used for MTT assay, and 6 teeth in TUNEL test. The Maxillary left and right, first and second molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group1 (immediately extraction), group 2 (cold preservation at 4$^{\circ}C$ for 1 week), group 3 (rapid cryopreservation in liquid nitrogen), group 4 (slow cryopreservation with magnetic field of 1 G), and group 5 (slow cryopreservation). F medium was used as preservation medium and 10% DMSO as cryoprotectant. After preservation and thawing, the MTT assay and TUNEL test were processed. One way ANOVA and Scheffe method were performed at the 95% level of confidence. The value of optical density obtained after MTT analysis was divided by the value of eosin staining for tissue volume standardization. In both MTT assay and TUNEL test, it had showed no significant difference among group 3, 4, and 5. And group 3 had showed higher viability of periodontal ligament cell than group 2. From this study, slow cryopreservation method with magnetic field can be used as one of cryopreservation methods.

Efficient Application of Westgard Multi-Rules and Quality Control Implementation Improvement (Westgard Multi-Rules의 효율적 적용과 조치사항의 개선)

  • Jung, Heung Soo;Oh, Youn Jung;Bae, Jin Soo;Baek, Jin Young;Hwang, Bo ra;Shin, Yong Hwan
    • The Korean Journal of Nuclear Medicine Technology
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    • v.21 no.1
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    • pp.60-64
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    • 2017
  • Purpose Westgard multi-rules application based on test quality improvement and commercialized international standard has been widely used in quality control. However, it is difficult to applicate the Westgard multi-rules in nuclear medicine in vitro tests due to the larger sample sizes and the simultaneous measurement of quality control material and patient sample. This study investigated the usefulness of Westgard multi-rules application in nuclear medicine in vitro tests. Materials and Methods A total of 282 systematic error multi-rules (22s, 101s) recorded in the samsung medical center computer system from January 2013 to June 2016 along with 117 cases of corrective measure record was analyzed. The Quality control implementation is recorded in Hospital information system were divided into 4 high-level areas including quality control material error, experimental procedural error, Kit lot number management error, and others. To prevent quality control material error, the existing method that each staff used their own method was changed. The staff who in charge of managing the quality control material was designated and daily consumption amount of every test was strictly controlled by one person. To prevent other errors, every test step was standardized so that the entire test procedures are identically implemented. Results The total quality control implementation was 117 cases; As a result, 62 quality control material errors were 62 cases, experimental process errors were 24 cases, Kit lot number control errors were 18 cases, and other errors were 13 cases. The quality control material error was corrected and could be used fresh materials within 2 days after thawing. The cases of systemic error were decreased to causes as quality control material error. The quality control materials were reduced above 10 vials to a monthly average. In addition, these errors of experimental processing and Kit lot number were improved by test standardization. Consequently, the cases of 101s and 22s in systematic error rules decreased at least 2 cases to a monthly average. Conclusion To confirm of systematic error through multi-rules application quickly, it is necessary to base on management of the QC material, target values and standard deviation. Moreover, in the event of a systematic error, it was found important to record measures based on test cause analysis. The experiment results are expected to contribute to internal quality control improvement and prompt and accurate result reporting through error recording and causal analysis based on Westgard multi-rules analysis.

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