This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.
To improve the efficiency of in vitro production of embryos with follicular oocytes in Korean Native cows, the recovery rates, in vitro maturation, fertilization and development, and the time required for collecting and processing oocytes by aspiration with or without slicing were evaluated comparatively. The ovaries were obtained from a local abattoir and placed in physiological saline at 25~28$^{\circ}C$ and brought to the laboratory within 3 hrs. The oocytes were collected by aspiration of follicles(2~6mm) with or without slicing ovaries after aspiration, and classified into Grade I, Grade II, Denuded, Expanded oocytes by the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules. Also the time required for each step of collecting and processing oocytes were measured. The cumulus cells were removed in some Grade I oocytes to measure their size and nuclear configuration before and after in vitro maturation. The Grade I oocytes were matured in vitro(IVM) for 24 hrs. in TGM-199 supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH, 1 $\mu$g /ml at 39$^{\circ}C$ under 5% C02 in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24hrs. and then the zygotes were cocultured in vitro (IVC) with bovine oviductal epithelial cells for 10 days. The results obtained were as follows: The number of oocytes recovered per ovary was averaged 6.6 by aspiration and 11.2 by slicing post aspiration, which summed to 17.8. The number of Grade I oocytes recovered per ovary was averaged 3.1 by aspiration and 3.6 by slicing, which summed to 6.7. The percentage of Grade I to total oocytes recovered was significantly(P<0.05) higher as 48.0 % in aspiration than 31.6% in slicing post aspiration. The time requlred for recovering a Grade I oocyte by aspiration and slicing was 1.1 and 2.5 min, respectively. The mean diameter of Grade I oocytes by aspiration and slicing was similar as 148.7 and 151.5$\mu$m, respectively. The percentage of Metaphase II stage oocytes after IVM for 24 hours was significantly (P
The incidence of ectopic pregnancy is increased since in vitro fertilization-embryo transfer (IVF-ET) has started. And because of the possibility of combined pregnancy after IVF-ET, bilateral adnexal area and other portion of pelvis should be examined carefully by transvaginal ultrasonography during follow-up period. The rate of combined pregnancy after IVF is estimated to be as high as 0.2~1%. Since the incidence of combined pregnancies is markedly higher with IVF, special attention to this possibility after ET is warranted. According to a recent report, the occurrence of interstitial pregnancy is estimated to be 1:3600 for all pregnancies achieved with IVF. The prevalence of interstitial pregnancies is 2~4% of ectopic pregnancies. Interstitial pregnancies tend to be diagnosed later than most other ectopic pregnancies, and if uterine rupture occurs, the resulting hemorrhage can be severe. Currently, about 2% to 3% of interstitial pregnancies are thought to carry a maternal death, which is twice that of other tubal gestations. We report a case of simultaneous left tubal and right interstitial pregnancies after IVF-ET with a brief review of literature.
Kim, Suh-Kyung;Kim, Young-Tae;Kim, Sun-Haeng;Rha, Jung-Ryul;Ku, Byung-Sahm
Clinical and Experimental Reproductive Medicine
/
v.17
no.2
/
pp.115-121
/
1990
Ultrasonically guided oocyte collection gradually replaces laparoscope in many IVF center. In present study, we compare the efficacy of both methods in our IVF program. Totally 377 cycles which were undertaken in vitro fertilization treatment were divided into 2 groups. Ultrasonically guided transvaginal follicular aspiration was performed in 188 cycles and laparoscopic follicular aspiration was performed in 189 cycles under local anesthesisa. The mean age for both groups was similar. Follicular recruitment was achieved with human menopausal gonadotropin (hMG) or a com bination of clomiphene citrate and hMG or a combination of FSH and hMG. In the ultrasonically guided aspiration group, 1821 follicles were aspirated with 61.8% of recovery rate (1125 oocytes), 81.5% of embryo transfer rate (145 cycles) and (17%), 26 cases intrauterine pregnancies were estabilished. In the laparoscopic group, 604 follicles were aspirated with 68.7% recovery rate (445 oocytes) and a 79.9% ET rate (127 cycles), 11 cases (8.7%) intrauterine pregnancies were estabilished. A valid comparison of these data is not possible because the 2 groups are dissimilar for factors known to influence oocyte development and recovery. No statistically significant differences could be demonstrated between 2 groups in all but the recovery rate and clinical pregnancy rate, In ultrasound group, the clinical pregnancy rate was significantly higher than that of laparoscope group. The potentially detrimental effect of CO2 pnemoperitonium present during laparoscope but not in ultrasound guided recovery on ova quality may underlie the observed difference in the clinical pregnancy rate between the 2 groups. Ultrasound guided aspiration seems to be as effective as laparoscopy in terms of oocyte retrieval and conception rate. Furthermore, the procedure is simple and inexpensive, it may replace laparoscopy as a method for oocyte collection in most patients who undergo IVF.
Proceedings of the Korean Society of Embryo Transfer Conference
/
2002.11a
/
pp.73-73
/
2002
Nuclear transfer (NT) techniques have advanced in the last years, and cloned animals have been produced by using somatic cells in several species including pig. However, it is difficult that the nuclear transfer porcine embryos development to blastocyst stage overcoming the cell block in vitro. Abnormal segregation of chromosomes in nuclear transferred embryos on genome activation stage bring about embryo degeneration, abnormal blastocyst, delayed and low embryo development. Thus, we are evaluated that the correlations of the frequency of embryo developmental rates and chromosome aberration in NT and In viかo fertilization (IVF) derived embryo. We are used for ear-skin-fibroblast cell in NT. If only karyotyping of embryonic cells are chromosomally abnormal, they may difficultly remain undetected. Then, we evaluate the chromosome aberrations, fluorescent in situ hybridization (FISH) with porcine chromosome 1 submetacentric specific DNA probe were excuted. In normal diploid cell nucleus, two hybridization signal was detected. In contrast, abnormal cell figured one or three over signals. The developmental rates of NT and IVF embryos were 55% vs 63%, 32% vs 33% and 13% vs 17% in 2 cell, 8 cell and blastocyst, respectively. When looking at the types of chromosome aberration, the detection of aneuploidy at Day 3 on the embryo culture. The percentage of chromosome aneuploidy of NT and IVF at 4-cell stage 40.0%, 31.3%, respectively. This result indicate that chromosomal abnormalities are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT associated with lower implantation rate, increase abortion rate and production of abnormal fetuses.
K. S. Chung;Ko, S. A;S. J. Song;J. T. Do;Park, Y. S.;Lee, H. T.
Korean Journal of Animal Reproduction
/
v.26
no.4
/
pp.385-394
/
2002
This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.
Bone marrow-derived cell (BMDC) therapy has numerous applications as potential biological cells for use in regenerative medicine. Here, we present an original case of endometrial atrophy associated with genital tuberculosis in a woman who achieved a live birth with BMDC. This 27-year-old woman came to our center with endometrial atrophy and primary infertility. She had a past history of genital tuberculosis and amenorrhea. Her husband's semen quality was normal. The patient was counseled for hysteroscopy due to thin endometrium and advised in vitro fertilization (IVF) with donor eggs in lieu of poor ovarian reserve. Several attempts of IVF with hormone replacement therapy (HRT) were made, but the desired thickness of the endometrium was not achieved. Uterine artery injection of BMDC through interventional radiology was given, followed by HRT for three months, which resulted in improved endometrium. This was subsequently followed by IVF with donor egg. The treatment resulted in the conception and delivery of a 3.1-kg baby boy through lower segment caesarean section with no antenatal, intranatal or postnatal complications. Recently, there has been massive interest in stem cells as a novel treatment method for regenerative medicine, and more specifically for the regeneration of human endometrium disorders like Asherman syndrome and thin endometrium, which was the reason behind using this strategy for treatment.
Objectives: The purpose of this study is to report a effect of herbal medicine at threatened abortion in the early stages of pregnancy. Methods: A 37-year-old female patient who had the abdomen pain with vaginal bleeding was taken Gyoaesamultang-gami and Antae-eum for 11 days at Dae-gu Haany University Hospital in Pohang. Results: During this treatment period, vaginal bleeding and abdomen pain were relieved. Also after all treatment, the effect had been continuing. Conclusions: Gyoaesamultang-gami is effective on threatened abortion. And the female patient deliver normally.
It has been suggested that the prognosis for fertility of the infertile patients with healed pelvic tuberculosis is very poor. Total 60 patients(77 cycles) with previous history of pelvic tuberculosis who underwent IVF-ET from January 1988 to March 1989 at SNUH were classified into three groups according to the principal histopathological lesions : tuberculous endometritis group(N=20, 28 cycles), tuberculous salpingitis group(N=32, 37 cycles) and pelvic peritoneal tuberculosis group(N=8, 12 cycles). To evaluate the effects of previous pelvic tuberculous lesions on ovarian follicular growth and development in controlled ovarian hyperstimulation for IVF-ET and its final outcome, serum E2 levels on the day of hCG administration(Day 0) and the day after hCG administration(Day +1), the number of ovarian follicles with mean diamete ${\geqq}$ 12 mm on Day 0, the number of oocytes retrieved by transvaginal aspiration, and pregnancy rate per cycle were measured and compared with control group(N=123, 161 cycles). There were no significant differences in cancellation rate during controlled ovarian hyperstimulation, total dosage of FSH and hMG administrated, menstrual cycle date(MCD) of hCG injection, serum E2 levels, the number of ovarian follicles with mean diameter ${\geqq}$ 15 mm, and the number of oocytes retrieved between pelvic tuberculosis group and control group. But in pelvic tuberculosis group, the number of ovarian follicles with mean diameter 12-14 mm, total number of ovarian follicles(${\geqq}$ 12 mm), and pregnancy rate per cycle were significantly decreased. These data suggest that previous pelvic tuberculous lesions have no significant adverse effects on the ovarian response to gonadotropin stimulation. IVF-ET proved to be an useful treatment modality for infertile patients with previous history of pelvic tuberculosis in spite of its relatively lowered pregnancy rate.
Objectives: This study aims to identify associated factors in female infertility treatment using herbal medicine, acupuncture and moxibustion. Methods: Acupuncture and moxibustion treatments were performed with intake of herbal medicines (Jokyungjongoktanggagam-bang, Sutaehwangagam-bang) for six menstrual cycles in 32 subfertile women. The variables selected for initial analysis were age, duration and type of infertility, frequency of previous In Vitro Fertilization (IVF) and live births, weekly frequency of sexual intercourse, number of treatment cycles, endometrial thickness on MCD15 and hormonal parameter (anti-mullerian hormone, follicle-stimulating hormone, luteinizing hormone, estrone). Results: Six patients became pregnant (18.75%) and seven patients withdrew. Factors influenced pregnancy were age (${\leq}35$ years), a shorter duration of infertility (<6 years), frequency of IVF (${\leq}3$), anti-mullerian hormone level (>1 ng/ml), and number of treatment cycles(${\leq}3$ cycles). Conclusions: Korean medicine is a useful and optimized treatment option for women with younger age, fewer previous IVF cycles, shorter infertile duration and normal anti-mullerian hormone level at first 3 cycles of treatment.
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