• Title/Summary/Keyword: Vitrified embryos

Search Result 99, Processing Time 0.022 seconds

Effects of the Oocyte and Developmental Stages of the Rat Embryos after the Vitrified Freezing on the Survival Rate(FDA-test) (Rat 난포란과 수정란의 발육단계별 유리화 동결 융해후 생존성(FDA-test)에 미치는 영향)

  • 고혁진;김중계
    • Journal of Embryo Transfer
    • /
    • v.11 no.1
    • /
    • pp.41-50
    • /
    • 1996
  • This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

  • PDF

Preferred strategy for euploid single embryo transfer in advanced maternal age: Fresh versus frozen

  • Fatma Ozdemir;Gokalp Oner;Semra Kahraman;Yucel Sahin;Hakan Yelke
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.51 no.1
    • /
    • pp.85-90
    • /
    • 2024
  • Objective: The purpose of this study was to compare fresh and frozen-thawed euploid blastocyst transfer protocols following preimplantation genetic screening (PGS) in cases of advanced maternal age. Methods: A total of 330 patients were examined retrospectively. PGS was performed on the embryos of 146 patients for whom fresh transfers were chosen. In contrast, frozen-thawed euploid single embryo transfer (ET) was selected after PGS for 184 patients, and their embryos were vitrified. The percentage of euploid embryos and rates of implantation, pregnancy, and pregnancy continuity, as well as clinical and biochemical abortion rates, were compared. Results: The numbers of retrieved oocytes, metaphase II oocytes, and fertilized ova were greater in the frozen-thawed group. The percentages of euploid embryos were comparable between the fresh and frozen-thawed groups (32% vs. 34.8%, respectively). The rates of implantation (46.6%vs. 62.5%), pregnancy (50% vs. 66.8%), ongoing pregnancy (38.4% vs. 53.8%), and live birth percentage (37.0% vs. 53.8%) were significantly higher in the frozen-thawed group. However, no significant differences were found in the clinical and biochemical abortion rates. Conclusion: The use of frozen-thawed single euploid ET is associated with increased implantation and pregnancy rates compared to fresh single euploid ET with PGS.

Effect of the Artificial Shrinkage on the Development of the Vitrified Bovine Embryos

  • Ha, A-Na;Cho, Su-Jin;Deb, Gautam-Kumar;Bang, Jae-Il;Kwon, Tae-Hyeon;Choi, Byeong-Hyun;Kong, Il-Keun
    • Journal of Embryo Transfer
    • /
    • v.25 no.1
    • /
    • pp.9-14
    • /
    • 2010
  • This study was conducted to find out the effects of artificial shrinkage (AS) on post-thaw development of bovine embryos. The blastocoelic cavity of blastocyst was punctured to remove its fluid contents and then incubated in the holding medium (HM) for 10 min. The punctured and non-punctured (control) blastocysts were equilibrated in vitrification solution 1 (VS1; TCM-199+20% FBS+10% EG) for 5 min and vitrification solution 2 (VS2; TCM199+20% FBS+35% EG+5% PVP+0.5 M Sucrose) for 1 min and vitrified by direct dropping into the liquid nitrogen. Vitrified blastocysts (punctured and control) were thawed and cultured in vitro (12 hr) for studying survival and hatching rates. The levels of shrinkage were measured by the volume of the blastocyst during equilibration in VS1 (at 1, 3 and 5 min of equilibration) and VS2 (at 30 and 60 sec of equilibration) that was considering the volume of non-punctured blastocyst in HM as 100%. The levels of shrinkage were higher in punctured group (62.4, 64.6, 64.3% at 1, 3 and 5 min in VS1; 50.6 and 52.7% at 30 and 60 sec in VS2) than control group (84.8, 86.6, 86.4% at 1, 3 and 5 min in VS1; 72.1 and 68.8% at 30 and 60 sec in VS2), but within each group the levels of shrinkage were similar. The survival (90.9%) and hatching (50.0%) rates of vitrified blastocysts at 12 hr post-thaw were higher in punctured group than that in control group (76.9% and 0.0% respectively). We confirmed that vitrification solutions (VS1 and VS2) have no toxic effect on the survival of blastocysts because the survival rates of blastocysts exposed to VS1 and VS2 for 24 hr were similar between punctured and control groups (94.3 vs. 96.0%; p>0.05). In conclusion, the preliminary data show that AS of blastocyst may improve survival and hatching rate after thawing.

In Vitro Development of Somatic Cell Nuclear Transferred Bovine Embryos Following Activation Timing in Mil Enucleated Oocytes Cryopreservation

  • Kim Seon-Gyun;Kim Eun-Yeong;Gil Gwang-S;Park Se-Yeong;Yun Ji-Yeon;Park Se-Pil;Jeong Gil-Saeng;Im Jin-Ho
    • Proceedings of the KSAR Conference
    • /
    • 2002.06a
    • /
    • pp.9-9
    • /
    • 2002
  • This study was to evaluate the in vitro survival of vitrified-thawed bovine MII enucleated (MIIe) oocytes according to activation timing and minimun volume cooling (MVC) method and their in vitro development after somatic cell nuclear transfer (SONT). Bovine oocytes were recovered from slaughtered bovine ovary and matured in TCM-199 supplemented with 10% FBS. (omitted)

  • PDF

Supplement of tauroursodeoxycholic acid in vitrification solution improves the development of mouse embryos

  • Lin, Tao;Lee, Jae-Eun;Shin, Hyun-Young;Oqani, Reza;Kim, So-Yeon;Jin, Dong-Il
    • Korean Journal of Agricultural Science
    • /
    • v.43 no.4
    • /
    • pp.575-580
    • /
    • 2016
  • This study was performed to determine whether supplementation of tauroursodeoxycholic acid (TUDCA), an endoplasmic reticulum (ER) stress inhibitor, during vitrified cryopreservation enhances the development of frozen mouse embryos. Mouse 8-cell stage embryos were collected and exposed to a cryoprotectant solution containing TUDCA or TM (tunicamycin, an ER stress inhibitor) at room temperature and stored in liquid nitrogen following vitrification. The final concentration of TUDCA or TM was $50{\mu}M$. The survival and development rates of mouse 8-cell stage embryos exposed to TUDCA- or TM-containing solutions at room temperature or stored in liquid nitrogen following vitrification were measured. There were no significant differences in survival rate and blastocyst formation rate among control, TUDCA, and TM groups after embryos were exposed to vitrification solutions at RT. When mouse 8-cell stage embryos were treated with TUDCA or TM and then stored in liquid nitrogen, the survival rates of control and TUDCA groups were significantly higher than for the TM group. Blastocyst formation rate of the TUDCA group following in vitro culture was significantly higher than that in control or TM groups. The TM group showed a lower (p < 0.05) blastocyst formation rate than the other two groups. Our results indicate that TUDCA supplementation during cryopreservation of mouse embryos could enhance their development capacity.

Clinical Application of Oocyte Cryopreservation I. Pregnancy and Delivery of Vitrified Human Oocytes in ART Program (난자동결보존의 임상적 응용 I. 유리화 난자동결 보존에 의한 임신과 분만)

  • 정형민;박이석;차광렬
    • Journal of Embryo Transfer
    • /
    • v.16 no.3
    • /
    • pp.245-250
    • /
    • 2001
  • This study was performed to evaluate whether vitrification method using ethyle glycol and eletron microscopic (EM) grid could be used far the cryopreservation of human oocytes in ART program. Surplus oocytes were obtained from consented IVF patients. These surplus human oocytes were frozen with our vitrification method, Oocytes were exposed to 1.5M ethylene glycol (EG) in DPBS far 2,5 minutes, followed by 5.5M EG plus 1.0M Sucrose in DPBS for 20 seconds. Then oocytes were transferred onto the EM grid and the grid was plunged into LN2 for storage. For thawing, oocytes containing EM grid were sequentially transferred in 1.0M, 0.5M, 0.25M, 0.125M and 0 M sucrose in DPBS solution at the intervals of 2.5 minutes. Thawed and survived oocytes were provided for ICSI. Embryos from vitrified oocytes were transferred to uterus of the patient on 4 to 5 days after ovulation in natural cycles of on 15 to 17 day of hormone replacement cycles. A total of 370 oocytes from 26 patients were thawed and 159 (43.0%) of them survived. One hundred thirty four oocytes (84.3%) were fertilized normally and 126 pre-embryos were transferred to 26 patients, resulting in 5 clinical pregnancies. The pregnancy rate per transfer was 19.2% and implantation rate was 4.0%. Among the five pregnant, 4 patients delivered 4 healthy babies and the one patient was 32-week ongoing pregnancy. From this results, vitrification using ethylene glycol as cryoprotectant and EM grid is a rapid and simple method that can be effectively applied for the cryopreservation of human oocytes in ART program.

  • PDF

Improvement of Straw Loading Method on Survival of Mouse IVF/IVC Blastocysts Cryopreserved by Vitrification (체외수정과 체외배양에서 생산된 생쥐 배반포기배의 초자화 동결에 있어서 Straw Loading 방법의 개선)

  • 김선의;엄상준;김은영;윤산현;박세필;임진호
    • Korean Journal of Animal Reproduction
    • /
    • v.20 no.1
    • /
    • pp.35-42
    • /
    • 1996
  • This study was carried out to investigate the effect of straw loading method and thawing protocol on the in vitro development of in vitro produced mouse blastocysts cryopreserved by vitrification. Three loading types of straw I, Il and III on loading and sealing method were made for vitrification. The ability of the solution on straw loading methods to remain vitreous during warming was tested by exposed in air for 1 to 10 s sec. and then plunged the vitrified straws into water bath at 25°C. Embryos to be vitrified were equilibrated to the 20% EG for 5min. and exposed in EFS 40 for 1min. The plug ends of Straw I and Straw II were sealed with straw powder and Straw III was treated straw powder, followed by heat sealing and then plunged into LN$_2$. The results obtained in these experiments were summarized as follows; 1) Straw I embryo column mostly changed from transparent to opaque upon thawing without exposure in air for 3-6 sec. Straw II embryo column was I improved partially but was not remained completely vitreous during warming. However, when Straw lll loading method was used, the embryo column was remained vitreous completely. 2) High survival rates and development rates of each groups (middle blastocysts and hatching blastocysts) of vitrified embryos were obtained by using Straw III loading method than Straw I method (P<0.05). And the range of s standard error was low in Straw lll method. 3) When the embryos vitrified-frozen were placed in air for 3, 5 and l0sec. and then warmed rapidly in water bath at 25$^{\circ}C$, the survival rates after 24h of culture were 72.7-87.1% and the development rates to hatching stage after 48h of culture were 34.0-48.4%. There were no significantly differences according to exposure time in air during warming. In conclusion, the present results showed that highly survival and low standard error of vitrified-frozen mouse bIastocysts were obtained by using straw lll loading, double sealing and appropriate 2 step warming method.

  • PDF

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
    • /
    • v.22 no.4
    • /
    • pp.349-357
    • /
    • 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).

  • PDF