• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.163-168
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• 1992

This study was carried out to set up the ovum bank for ovum donation and to determine the best freezing method for human immature oocytes. Human immature follicular oocytes were cryopreserved by slow freezing and rapid thawing method. Immature follicular oocytes were treated by propanediol(PROH) solution by 2 and 4 step method in protocols A & B, respectively. In protocol C, immature oocytes were exposed to sucrose prior to treatment of PROH by 4 step method. We compared survival rate, maturation rate, and fertilization rate of immature oocytes among three protocols. Results were as follows. 1. Oocytes treated by the protocol C showed the highest survival rate( 70.3 %) and maturation rate(34.6%) after thawing. 2. Survival rate of oocytes treated by the protocol C was significantly higher than that of the protocol B after thawing(p<0.05). In conclusion, treatment of oocytes with sucrose prior to expose PROH was the best freezing method. Sucrose may have reduced the toxic effect of cryoprotectant to oocytes. We failed to induce fertilization of oocytes, which were treated by any protocols, by conventional insemination method, but obtained 28.8% fertilization rate by using partial zona dissection(PZD) method. This result suggests that micromanipulation(PZD) of the thawed oocytes before insemination will improve the fertilization rate.

• 한국산학기술학회논문지
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• 제16권11호
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• pp.7555-7563
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• 2015

이온강도 0.10 M, $25^{\circ}C$ 수용액에서 Zn(II)이온과 2-(2-hydroxyethylamino)-2-(hydroxymethyl)-1,3-propa nediol(Monotris), Bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane(Bistris)과의 착물 형성을 전위차법으로 연구하였다. Zn(II)-Monotris 시스템에서 Monotris(L) 착물 $ZnL^{2+}$는 리간드의 히드록실 산소 원자 중에서 한 개의 산소 원자 뿐만 아니라 아민 질소가 배위되었다. 배위되어 있던 히드록실기의 수소 이온이 염기성 영역에서 해리되었다. Zn(II)-Bistris 시스템에서 Bistris(L) 착물 $ZnL^{2+}$는 리간드의 히드록실 산소 원자 중에서 두 개의 산소 원자 뿐만 아니라 아민 질소가 배위되었다. 배위되어 있던 히드록실기들 중 한 개의 히드록실기에서 수소 이온이 염기성 영역에서 해리되었다. 추가적으로 다른 한 개의 히드록실기에서 수소이온이 강염기 영역에서 해리되었다. $ZnL^{2+}$, $ZnLH_{-1}{^+}$, $ZnLH_{-2}$, $Zn_2L_2H_{-2}{^{2+}}$ 그리고 $Zn_2L_2H_{-3}{^+}$의 평형 상수값을 계산하였다.

• 청정기술
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• 제25권2호
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• pp.114-122
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• 2019

최근 재생자원으로부터 바이오 폴리올을 합성하는 것이 주목을 받고 있다. 특히 고분자의 합성에서 이러한 바이오 폴리올을 활용하는 것은 대단히 중요한 과제이다. 폴리카보네이트 폴리올/바이오 폴리올(PO3G: 옥수수 당의 발효에 의해 제조된 1,3-프로판 디올로부터 제조된 폴리트리메틸렌 에터 글리콜), 메틸렌디페닐디이소시아네이트 및 1,4-부탄디올을 사용하여 일련의 디메틸포름아미드(DMF) 기반 폴리우레탄을 합성하였다. 본 연구에서는 폴리우레탄 필름의 특성과 습식 인조피혁의 셀(cell) 특성을 조사하였다. 폴리카보네이트 폴리올/바이오 폴리올에서 바이오 폴리올의 함량이 증가할수록 폴리우레탄 필름의 인장강도는 감소하지만 연신율은 증가하였으며, 유리전이온도는 낮아짐을 알 수 있었다. 또한 습식공법에 의한 인조피혁 단면을 분석한 결과 폴리카보네이트 폴리올 함량이 증가함에 따라 인조피혁에 형성된 셀의 수와 균일성이 증가함을 알 수 있었다. 이 결과로부터 적정량의 바이오 폴리올을 사용한 DMF 기반 폴리우레탄의 경우에 충분히 인조피혁에 사용할 수 있음을 알 수 있었다. 바이오 탄소 함량은 폴리우레탄의 제조에 사용한 바이오 폴리올의 함량 증가에 따라 비례하여 증가하였다.

• KSBB Journal
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• 제9권3호
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• pp.246-252
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• 1994

유기용매 THF에서 bis(2, 2, 2-trichloroethyl) glutarate와 diol을 기질로 하여 PPL에 의한 tran sesterification 반응을 통해 polyester를 합성하였다. 1. 서로 다른 source의 lipase로 이 반응을 시켜본 결과 PPL이 가장 반응을 잘 진행시켰으며 Humi cola lanuginos와 Pseudomonas sp.의 Ii pase들도 어느 정도 반응을 진행사켰으며 Rhizopus의 lipase들 은 반응을 전혀 진행시키지 못했다. 2. Diol로 ethyleneglycol, 1, 3~propanediol, 1,4­b butanedi이은 모두 거의 비슷한 정도로 반응이 진행 되었으며 secondary alcohol인 glycerol의 경우 이들에 비해 반응이 매우 느리게 진행되었다. 이는 glycer이 에 의 해 효소가 steric hinderance를 받기 때문인 것으로 추정된다. 3. Diol로 여러 가지 분자량의 PEG(M = 300-1000)를 사용한 결과 PEG-400이 가장 잘 반응되었으며 이를 제외하고는 분자량이 클수록, 즉 chain의 길이가 갈수록 반응속도가 느렸다. 그리고 이렇게 chain의 길이가 비교적 긴($C_{12}-C_{45}$) PEG플 diol 로 사용했을 때는 monotransesterication 반응이 끝 난 이후에는 반응이 거의 더 이상 진행되지 않았다. 4. 유기용매로는 비교적 hydrophilic한 THF, ether, acetonitrile 등에 셔 잘 반응되었다. 5. $20^{\circ}C$ 에서 $60^{\circ}C$의 온도범위에서 반응온도가 높 을수록 반응속도가 빨랐으며 184시간 반응된 반응 생 생물의 평균분자량은 모두 2200←2300 daltons로 반응온도의 영향을 받지 않는 것으로 관찰되었다. 8. NMR에 의한 end group analysis를 이용하여 반응 생성물을 분석한 결과 수평균분자량은 1500­4 4000 daltons이었다.

• 청정기술
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• 제25권1호
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• pp.7-13
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• 2019

일련의 dimethylformamide (DMF) 기반 폴리우레탄은 메틸렌 디 페닐 디 이소시아네이트(MDI) 1,4-부탄 디올 및 바이오 폴리올(1,3-프로판 디올: B-POL)에 기초한 폴리 트리 메틸렌 에테르 글리콜/폴리 에스터 폴리올(1,4-부탄디올: H-PET)으로 부터 합성하였다. 본 연구에서는 바이오 폴리올(B-POL)/폴리에스터 폴리올(H-PET)의 조성이 폴리우레탄의 물성에 미치는 영향을 조사하였다. B-POL/H-PET 혼합물의 B-POL 함량이 증가할수록 폴리우레탄의 소프트 세그먼트의 유리전이온도(Tgs)와 인장 강도는 감소하는 반면, 파괴 신도 및 인열 강도는 증가하였다. 한편 합성된 DMF 기반 폴리우레탄을 이용하여 습식공정으로 인조피혁을 제조하였다. 인조피혁에 형성된 다공성 셀의 평균 크기 및 밀도(단위부피당 셀의 수)에 미치는 B-POL/H-PET 조성의 영향의 차이는 거의 없는 것을 알 수 있었다. 이러한 결과들로부터 바이오 폴리올 기반 폴리우레탄을 습식공정으로 제조되는 인조피혁용으로 사용하는데 별 문제가 없음을 알 수 있었다.

• Clinical and Experimental Reproductive Medicine
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• 제31권1호
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• pp.59-65
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• 2004

Objectives: The aim of this study was to assess toxicities of cryoprotectants. Methods: Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and 1, 2-propanediol (PROH), were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or PROH. Embryo development was evaluated up to the blastocyst stage. Blastocysts were stained with bis-benzimide to evaluate the cell count and with terminal deoxynucleotidyl transferase mediated dUTP nick labeling (TUNEL) to assess apoptosis. Results: The total cell count of blastocysts that were treated with DMSO at the 2-cell stage was significantly lower than that were treated with PROH ($75.9{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the PROH treated group (p<0.05). Both DMSO ($14.2{\pm}1.5$) and PROH ($11.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.2{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the PROH treated group (p<0.001). Conclusions: The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or PROH at room temperature. When comparing two cryoprotective agents, PROH appeared to be less toxic than DMSO at least in a murine embryo model.

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.111-118
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• 1997

Intracytoplasmic sperm injection (ICSI) recently has been utilized widely as the most successful technique to overcome the unfertilization problem in cases of severe male infertility in couples who could not be treated by conventional IVF. Recently, indications of ICSI have been extended further and more fertilized oocytes become available. Thus, it is necessary to examine the efficiency of freezing the surplus embryos obtained from ICSI. We compared the survival rate and the future outcome of cryopreserved embryos obtained either after conventional IVF or ICSI during the same period. After ICSI or IVF, five best-quality embryos from each patient were transferred in the stimulation cycle and the surplus pronuclear (PN) stage oocytes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propanediol (PROH) as a cryoprotectant. A total of 792 embryos from ICSI trial were thawed and 65.2% (516/792) survived. The survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 63.5%, 68.2%, 64.0%, respectively. After 111 transfers, 34 pregnancies were achieved, corresponding to a clinical pregnancy rate of 30.6% per transfers. We thawed 1033 embryos from IVF trials and 57.5% (594/1033) survived. In IVF cycle, the survival rates of PN stage oocyte, multicellular embryo and PN + multicellular embryo were 58.2%, 65.2%, 40.2%, respectively. Thirty eight clinical pregnancies were established after 134 transfers, corresponding to a pregnancy rate of 28.4% per transfer. The cleavage rate of thawed PN stage oocytes from ICSI trial (61.3%) was significantly higher than those from conventional IVF (53.4%). The developmental rates of good embryo (${\geqq}$ grade II) in thawed PN stage oocytes obtained from conventional IVF and ICSI were 63% and 65%, respectively. We concluded that PN stage oocytes, multicellular embryos resulting from ICSI procedure can be successfully frozen/thawed with reasonable clinical pregnancy rates comparable to those of IVF.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.59-65
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• 2000

Objective: The purpose of this study was to determine the important factors affecting survival and pregnancy rate in frozen-thawed embryo transfer cycles. Methods: we performed retrospective analysis in 738 cycles of frozen-thawed embryo transfers, in relation to the insemination methods, the freezing stage of embryo, patient's age, infertility factors and the origin of injected sperm in ICSI cycles. After conventional IVF or ICSI, the supernumerary PN stage zygotes or multicellular embryos were cryopreserved by slow freezing protocol with 1,2-propanediol (PROH) as a cryoprotectant. Results: The survival rates of thawed embryos were 69.3% (1585/2287) in conventional IVF group and 71.7% (1645/2295) in ICSI group. After frozen-thawed embryo transfers, 27.0% (92/341) and 32.0% (109/341) of pregnancy rates were achieved in conventional IVF and ICSI group, respectively. There were no significant difference in the survival and pregnancy rates according to the insemination methods, the freezing stage and patient's age. However, the pregnancy rate (36.2%) of male factor infertility was significantly higher than the tubal (27.2%) and other female factor infertility (22.9%). In ICSI group, the origin of injected sperm did not affect the outcome of frozen-thawed embryo transfer cycles. Conclusion: The present study demonstrates that acceptable clinical outcomes can be achieved after the transfer of frozen-thawed embryos regardless of the stage of embryos for freezing, the patient's age and the origin of injected sperm.

• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.281-292
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• 1997

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.