• Clinical and Experimental Reproductive Medicine
• /
• 제34권4호
• /
• pp.253-273
• /
• 2007

목 적: 본 연구는 연구자로 하여금 [잔여배아보관실적대장]과 [잔여배아제공실적대장]을 명확하게 작성토록 하고 관리자 및 감독자로 하여금 이를 합리적이고 통일된 관리 및 점검을 할 수 있도록 하기 위한 방법을 찾고자 한 것이었다. 연구방법: 1994년부터 2004년까지 44호 기관에 보관되어 있는 자료들을 근거로 연도별 [배아의 생성 및 이용에 관한 해당연도 현황], [연도별 잔여배아의 냉동보관과 해당연도 해동현황], [냉동배아의 해동 및 이용에 관한 해당연도 현황], [냉동배아의 제공 및 인수에 관한 해당연도 현황], [해당연도 냉동배아폐기대장], [냉동배아의 관리 및 이용에 관한 해당연도총괄대장]을 도안하고 작성하였다. 결 과: 1994년부터 2004년까지 연도별 [44호 배아의 생성 및 이용에 관한 해당연도 현황], [44호 연도별 잔여배아의 냉동보관과 해당연도 해동현황], [44호 냉동배아의 해동 및 이용에 관한 해당연도 현황], [44호 냉동배아의 제공 및 인수에 관한 해당연도 현황], [44호 해당연도 냉동배아폐기대장], [44호 냉동배아의 관리 및 이용에 관한 해당연도 총괄대장]을 구축하였던 바 해당항목들이 상호일치하고 있었다. 결 론: 본 연구에서 얻어진 결과들은 연도별 [해당기관 해당연도 잔여배아보관실적대장]과 [해당기관 해당연도 잔여배아제공실적대장]을 작성하는 기초자료로서 뿐만 아니라 [배아생성의료기관에 보관해야 할 참고문서]로도 충분하다고 사료된다.

• International Journal of Oral Biology
• /
• 제38권4호
• /
• pp.161-167
• /
• 2013

Human dental pulp stem cells (DPSCs) are multi-potent mesenchymal stem cells that have several differentiation potentials. An understanding of thetissues that differentiate from these cells can provide insights for future regenerative therapeutics and tissue engineering strategies. The mesiodens is the most frequent form of supernumerary tooth from which DPSCs can differentiate into several lineages similar to cells from normal deciduous teeth. Recently, it has been shown that nanoscale structures can affect stem cell differentiation. In our presentstudy, we investigated the effects of a 250-nm nanoscale ridge/groove pattern array on the osteogenic and adipogenic differentiation of dental pulp cells from mesiodenscontaining human DPSCs. To this end, the expression of lineage specific markers after differentiation induction was analyzed by lineage specific staining and RT-PCR. The nanoscale pattern arrayed surface showed apositive effect on the adipogenic differentiation of DPSCs. There was no difference between nanoscale pattern arrayed surface and conventional surface groups onosteogenic differentiation. In conclusion, the nanoscale ridge/groove pattern arrayed surface can be used to enhance the adipogenic differentiation of DPSCs derived from mesiodens. This finding provides an improved understanding of the effects of topography on cell differentiation as well as the potential use of supernumerary tooth in regenerative dental medicine.

• Clinical and Experimental Reproductive Medicine
• /
• 제43권3호
• /
• pp.181-184
• /
• 2016

The aim was to report a healthy live birth using re-vitrified-warmed cleavage-stage embryos derived from supernumerary warmed embryos after frozen embryo transfer (ET) in a patient with recurrent implantation failure (RIF). The case was a 39-year-old female with a history of polycystic ovarian syndrome and adenomyosis, along with RIF. After ovarian hyperstimulation, 33 cumulus-oocyte complexes were retrieved and fertilized with conventional in vitro fertilization and intracytoplasmic sperm injection. Because of the risk of ovarian hyperstimulation syndrome, 16 grade B and C embryos were vitrified. After 3 and 6 months, 3 and 4 B-C warmed embryos were transferred to the uterus, respectively. However, implantation did not take place. Ten months later, four embryos were warmed, two grade B 8-cell embryos were transferred, and two embryos were re-vitrified. One year later, the two re-vitrified cleavage-stage embryos were warmed, which resulted in a successful live birth. This finding showed that following first warming, it is feasible to refreeze supernumerary warmed embryos for subsequent ET in patients with a history of RIF.

• Clinical and Experimental Reproductive Medicine
• /
• 제27권1호
• /
• pp.59-65
• /
• 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
• /
• 제24권1호
• /
• pp.111-118
• /
• 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.

• 한국가축번식학회지
• /
• 제22권1호
• /
• pp.11-17
• /
• 1998

본 연구는 생쥐 완전탈출 배반포기배의 체내 발달율을 조사하기 위해 실시하였다. 공시된 완전탈출 배반포기배는 체내에서 생산된 전핵기 수정란을 5일과 6일동안 체외배양하여 얻었으며, 완전탈출 배반포기배의 직경을 기준으로 small(S-HBs), medium(M-HBs), large(L-HBs)로 구분하였다. 그 결과를 요약하면 다음과 같다. 1) 체외배양 4일째에 얻어진 배반포기배를 $24\sim48$시간동안 추가배양했던 바, 배양 5일과 6일째에 완전탈출 배반포기배의 발달율은 29.1%와 22.8%였다. 2) 또한, 완전탈출 배반포기배의 총 세포수를 조사하였던 바, S-HBs ($77.7\pm5.3$, $59.6\pm4.4$), M-HBs ($83.7\pm4.0$, $66.8\pm3.5$), L-HBs ($100.7\pm2.6$, $88.9\pm3.8$)로 나타나, 완전탈출 배반포기배의 크기가 증가함에 따라 총 세포수도 증가한다는 것을 알 수 있었다. 특히, S-HBs와 L-HBs의 총 세포수간에는 유의적인 차이가 있었다.(p<0.01). 3) 분류된 완전탈출 배반포기배를 가임신 3일된 대리모의 자궁에 이식하였을 때, 배양 5일째의 임신율과 착상율이 S-HBs (28.6%, 15.7%), M-HBs (44.4%, 30.9%), L-HBs (62.5%, 49.1%)로서 완전탈출 배반포기배의 크기가 커질수록 증가하였던 반면, 배양 6일째에는 이러한 양상을 볼 수 없었다. 그러나, 전체 착상율에 대한 정상산자율을 조사하였던 바, 배양 5일째의 S-HBs (87.5%)가 다른 군에 비하여 유의하게 높게 나타났다. (p<0.01). 따라서, 이러한 결과로 미루어 볼 때, 체외에서 배양된 건강한 완전탈출 배반포기배는 높은 임신율과 착상율 및 정상산자율을 얻을 수 있음을 확인하였던 바, 이는 인간포배기 배아이식시 잉여의 완전탈출 배반포기배의 유용성 검토를 위한 기초자료로서 이용될 수 있음을 시사한다고 하겠다.

• Clinical and Experimental Reproductive Medicine
• /
• 제33권4호
• /
• pp.237-243
• /
• 2006

목 적: 동결-융해 배아이식은 보조생식술에서 환자들에게 보다 많은 임신의 기회를 제공해줄 수 있는 방법으로 이용되고 있다. 본 연구에서는 예후가 좋지 않은 환자들에서 동결-융해 배아이식의 효용성을 알아보고자 하였다. 연구방법: 나이가 많은 고령 환자군 (38~44세)과 신선주기 배아이식에서 임신 실패군을 연구대상으로 하였다. 과배란 유도를 통해 채취한 난자를 일반적인 체외수정 또는 세포질내 정자주입술을 시행하여 수정을 유도하고, 잉여의 전핵 또는 난할 시기의 배아를 완만동결법으로 동결하였다. 동결보관 배아는 급속융해법으로 융해하여 호르몬요법을 시행한 환자의 자궁에 이식하였다. 신선 배아이식과 동결-융해 배아이식 과정에서의 배아 상태, 임신율, 착상률 등을 통계적인 방법으로 분석하였다. 결 과: 나이가 많은 고령군에서 신선 배아이식을 시행한 환자들과 동결-융해 배아이식을 시행한 환자들의 평균 연령은 $40.0{\pm}1.8$세 (n=206)와 $39.9{\pm}1.9$세 (n=69)로 통계적으로 유의한 차이가 없었으나, 임상적인 임신율과 착상률은 동결-융해 배아이식에서 29.0%와 11.2%로 신선 배아이식의 16.5%와 7.0%에 비해 통계적으로 유의하게 높게 나타났다. (p<0.05). 첫 번째 신선 배아이식에서 임신 실패군의 연속되는 신선 배아이식 환자군 ($31.2{\pm}2.3$, n=40)과 동결-융해 배아이식 환자군 ($31.9{\pm}3.1$, n=119)에서의 평균 연령은 차이가 없었으며, 임상적 임신율 (42.5% vs 40.3%)과 착상률 (22.6% vs 18.8%)도 유사하였다. 결 론: 본 연구에서는 동결-융해 배아이식이 고령 환자들에서 효과적으로 임신율과 착상률을 높일 수 있음을 보여주고 있다. 이러한 결과는 과배란 유도에 따른 자궁의 착상 환경 변화가 고령 환자들에서 임신율과 착상률을 저하시키는 것과 관련이 있을 것으로 생각된다.

• Clinical and Experimental Reproductive Medicine
• /
• 제27권1호
• /
• pp.67-74
• /
• 2000

Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.