• Title/Summary/Keyword: Reproductive Techniques

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Effect of Various Caffeine Concentrations and Fertilization Time in In Vitro Fertilization of Canine Oocytes

  • Kim, Bong-Soo;Yoo, Dong-Hoon;Shin, Mi-Jung;Kim, Hae-Jung;Lee, Dong-Seok;Hyun, Byung-Hwa;Lee, Sang-Gyu;Park, Young-Sik;Ha, Ji-Hong;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.217-222
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    • 2008
  • The techniques of IVM, IVF and IVC of canine oocytes may provide useful information for gamete salvage programs and the conservation of endangered canidae. This investigation has been made to determine the efficiency of in vitro maturation (IVM) as a basic experiment to study the development of canine oocytes after in vitro fertilization (IVF). The rate of oocytes developing to the Mil stage was higher in the hormone treated group (10 IU/ml hCG+eCG, 14.7%, p<0.05) than in the control group (0 IU/ml hCG+eCG, 10.0%). The monospermy and pronuclear rates of canine oocytes were investigated after caffeine treatment on IVF. Canine oocytes were fertilized in the Fert-TALP medium supplemented with 0, 10, 20 or 30 mM caffeine (Fert I, Fert II, Fert III or Fert IV, respectively). The highest pronuclear formation rate was obtained in the Fert I for 24 h IVF (6.7%, 6/89). Therefore, it is believed that unlike in other mammals, caffeine in canine IVF does not increase the efficiency of fertilization rate, and is not an important factor.

The G-, C-, and NOR-Banded Karyotypes of Korean Short-hair Cat(Felis catus) (G-, C-Banding 및 NORs 분포 양상에 따른 한국산 단모고양이(Felis catus)의 핵형분석)

  • Cho, Eun Jung;Kong, Il Keun;Sohn, Sea Hwan
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.161-167
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    • 2013
  • The karyotype of Korean short-hair cat was presented using the G-, C- and NOR-banding techniques. For chromosomes preparation, the fetus skin fibroblast cells were cultured and metaphases were obtained. In results, the Korean short-hair cat had 38 chromosomes with XX or XY, which consisted of 5 pairs of metacentric chromosomes(Group A and C), 3 pairs of submetacentric chromosomes (Group B), 6 pairs of medium metacentric chromosomes except for 1 pair of medium submetacentric D2 chromosomes (Group D, E), 2 pairs of acrocentric chromosomes(Group F) and metacentric X and Y sex chromosomes. In G-banding analysis, the Korean short-hair cat exhibited a typical and identical G-banding pattern in each homologous chromosome. Total number of bands and landmarks on the G-banded chromosomes of Korean short-hair cat well correspond to those of international standardization of karyotype of domestic cat. The heterochromatins of Korean short-hair cat chromosomes distributed at terminal and/or centromere regions on almost chromosomes by C-banding analysis. In addition, the C-banding pattern showed greatly heteromorphic in some chromosomes. Using the AgNOR-staining, we found the nucleolar organizer regions(NORs) of Korean short-hair cat located at chromosomes 1p12 site in E group. The quantity and number of NORs were constant among cells.

Comparison of In Vitro Development of Porcine Embryos Derived from Transfer of Embryonic Germ Cell Nuclei into Oocytes by Electrofusion and Piezo-Driven Microinjection

  • Ahn, Kwang-Sung;Won, Ji-Young;Heo, Soon-Young;Kang, Jee-Hyun;Shim, Ho-Sup
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.127-131
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    • 2007
  • Embryonic germ (EG) cells are undifferentiated stern cells isolated from cultured primordial germ cells (PGC). These cells share many characteristics with embryonic stem cells including morphology and pluripotency. Undifferentiated porcine EG cell lines demonstrating capacities of differentiation both in vitro and in vivo have been established. Since EG cells can be cultured indefinitely in an undifferentiated state, whereas somatic cells in primary culture are often unstable and have limited lifespan, EG cells may provide inexhaustible source of karyoplasts in nuclear transfer (NT). In this study the efficiencies of NT using porcine EG and fetal fibroblast cells were compared. Two different techniques were used to perform NT. With conventional NT procedure (Roslin method) involving fusion of donor cells with enucleated oocytes, the rates of development to the blastocyst stage in EG and somatic cell NT were 16.8% (59/351) and 14.5% (98/677), respectively. In piezo-driven microinjection (Honolulu method) of donor nuclei into enucleated oocytes, the rates of blastocyst formation in EG and somatic cell NT were 11.9% (15/126) and 9.4% (9/96), respectively. Regardless of NT methods used in this study, EG cell NT gave rise to comparable rate of blastocyst development to somatic cell NT. Overall, EG cells can be used as karyoplast donor in NT procedure, and embryos can be produced by EG cell NT that may be used as an alternative to conventional somatic cell NT.

Effective Application of Multiplex RT-PCR for Characterization of Human Embryonic Stem Cells/ Induced Pluripotent Stem Cells (다중 역전사 중합효소 연쇄 반응(Multiplex RT-PCR)을 이용한 인간배아 줄기세포 및 유도만능 줄기세포의 효과적인 분화 양상 조사)

  • Kim, Jung-Mo;Cho, Youn-Jeong;Son, On-Ju;Hong, Ki-Sung;Chung, Hyung-Min
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.1-8
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    • 2011
  • Techniques to evaluate gene expression profiling, such as sufficiently sensitive cDNA microarrays or real-time quantitative PCR, are efficient methods for monitoring human pluripotent stem cell (hESC/iPSC) cultures. However, most of these high-throughput tests have a limited use due to high cost, extended turn-around time, and the involvement of highly specialized technical expertise. Hence, there is an urgency of rapid, cost-effective, robust, yet sensitive method development for routine screening of hESCs/hiPSCs. A critical requirement in hESC/hiPSC cultures is to maintain a uniform undifferentiated state and to determine their differentiation capacity by showing the expression of gene markers representing all three germ layers, including ectoderm, mesoderm, and endoderm. To quantify the modulation of gene expression in hESCs/hiPSC during their propagation, expansion, and differentiation via embryoid body (EB) formation, we developed a simple, rapid, inexpensive, and definitive multimarker, semiquantitative multiplex RT-PCR platform technology. Among the 9 gene primers tested, 5 were pluripotent markers comprising set 1, and 3 lineage-specific markers were combined as set 2, respectively. We found that these 2 sets were not only effective in determining the relative differentiation in hESCs/hiPSCs, but were easily reproducible. In this study, we used the hES/hiPS cell lines to standardize the technique. This multiplex RT-PCR assay is flexible and, by selecting appropriate reporter genes, can be designed for characterization of different hESC/hiPSC lines during routine maintenance and directed differentiation.

Development of Vitrified Bovine Oocytes following Intracytoplasmic Sperm Injection (ICSI)

  • Yeo, H-J;Ock, S-A;Yea, E-H;Lee, H-J;Choe, S-Y;Park, G-J
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.6-6
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    • 2001
  • Oocyte freezing has become a prevalent source for related reproductive technologies. This study was carried out to evaluate viability of post-thawed bovine oocyte injected DTT-treated sperm following by two different activation stimuli (Group 1, 5 M ionomycin, 5 min + CR1aa, 3 h . 1.9 mM dimetylaminopurine (DMP), 3 h; group 2, ionomycin + 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CHX), 5h). The techniques of ultra-rapid freezing used in this study were essentially similar to those of described by Vajta et al (Theriogenology 1999; 52:939-948), Denuded oocytes at 22 h of culture were exposed to cryoprotectant (3.2 M Ethylene glycol, 2.36 M DMSO, 0.6 M sucrose), and followed by freezing in electron microscopic grid. After thawing the oocytes were transferred back into the drop of maturation medium and cultured for additional 2 h before being subjected to ICSI. All eggs were then cultured in CRlaa medium, and transferred into M199+10% FCS on day 4. The culture was maintained until day 9. In Experiment 1, frozen-ICSI eggs were compared on development into blastocyst to those of unfrozen and IVF control. Those eggs were activated with the method of group 2. A higher proportion of unfrozen-ICSI and IVF eggs developed into cleavage and blastocysts than of frozen-ICSI eggs (65% and 13%; 71% and 23% vs. 39% and 8%; P<0.05). In Experiment 2, development and ploidy of embryos made from group 1 were compared to those from group 2. Between groups there did not differ on the rates of development, however, chromosomal abnormality in group 1 was significantly higher than in group 2 (49% vs. 30%; P<0.05). The present result suggests that frozen bovine oocytes can be used for ICSI.

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Cervical Cancer Prevention Knowledge and Attitudes among Female University Students and Hospital Staff in Iran

  • Asgarlou, Zoleykha;Tehrani, Sepideh;Asghari, Elnaz;Arzanlou, Mohammad;Naghavi-Behzad, Mohammad;Piri, Reza;Sheyklo, Sepideh Gareh;Moosavi, Ahmad
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4921-4927
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    • 2016
  • Background: Cervical cancer is a major preventable cancers. The, current study aimed to assess relevant knowledge and attitude of female students and hospital staff in Iran. Method: This cross-sectional study was conducted in Medical and Nursing faculties and hospitals of East-Azerbaijan Province of Iran. Participants were medical and paramedical female students and female staff in hospitals selected by stratified random sampling techniques. Tools for data collection were questionnaires for which validity and reliability had been verified (${\alpha}=0.8$). Descriptive and inferential statistics were used to analyze data with SPSS.16. Result: Response rates were 71 % (426 from 600) and 63.5% (254 from 400) for students and staff, respectively. Some 29.1% admitted that they had no information about cervical cancer, only 70 (10.3%) thinking their knowledge as high, 360 (52.9%) as intermediate, and 237 (34.9%) as low. While 93% of participants considered cervical cancer as a severe health problem, the only statistically significant relationships with knowledge were for education (p<.001) and occupation (p<.001) variables. Conclusion: Given the importance of the roles of medical students and personnel as information sources and leaders in health and preventive behavior, increasing and improving their scientific understanding seems vital. Comprehensive and appropriate education of all people and especially students and personnel of medical sciences and improving attitudes towards cervical cancer and its monitoring are to be recommended.

Immunologic Aspects at the Feto-Maternal Interface (태아모체간 계면에서의 면역학적 측면)

  • 정인배
    • Development and Reproduction
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    • v.5 no.2
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    • pp.93-100
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    • 2001
  • Precise mechanism by which the fetus can escape from mother's immune rejection is not well understood yet over the last 50 years. The clarification of immune mechanism at the feto-maternal interface is very important, because this can be a common pathogenesis of various pathologic conditions including spontaneous abortion, habitual abortion fetal growth restriction preeclampsia, implantation failure after assisted reproductive techniques, and fetal death. In this review, current hypothetical contents were described with the priority of importance: 1) The center of this mechanism is cross-talk between the expression of HLA-C, E, G on the extravillous cytotrophoblasts and their receptors on decidual NK cell, 2) immunomodulation, 3) innate immunity is the main immunologic mechanism, 4) various mechanisms besides HLA system(eq. complement) may be associated. The overall balance of immunomodulation among these mechanisms should result in the outcome of each pregnancy. Further researches regarding the regulation of HLA system, roles of cytokines, complements should be followed in the future.

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Application of Embryo Transfer Technology (수정란 이식 기술의 응용)

  • Lim, Hyun-Joo;Son, Jun-Kyu;Yoon, Ho-Beak;Baek, Kwang-Soo;Choe, Chang-Yong;Kim, Sidong;Kwon, Eung-Gi
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.163-168
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    • 2013
  • Embryo transfer (ET) technology is of high importance in modern cattle breeding programs. ET is one step in the process of removing one or more embryos from the reproductive tract of an outstanding donor female and transferring them to one or more recipient females. Embryos also can be produced in the laboratory via techniques such as in vitro fertilization (IVF). But the actual transfer of an embryo is only one step in a series of processes that may include some or all of the following: superovulation and insemination of donors, collection of embryos, isolation, evaluation and short-term storage of embryos, micromanipulation and genetic testing of embryos, freezing of embryos and embryo transfer. Cryopreservation and direct transfer of frozen-thawed embryos is common-place with pregnancy rates near that of fresh embryos. Polymerase chain reaction (PCR) technology is currently being used for sexing embryos, and this technology will be used for "embryo diagnostics" and "embryo genomics" in the future. Although, many limitations and problems remain to overcome, these and other new technologies promise to change livestock breeding drastically in the next decade.

Application of Mini-abdominoplasty after Conservative Excision of Extensive Cesarean Scar Endometriosis

  • Lee, Eui Tai;Park, Hyun Min;Lee, Dong Geun;Shin, Kyung Jin;Kim, Hak Soon;Sung, Ro Hyun;Ryu, Dong Hee
    • Archives of Plastic Surgery
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    • v.39 no.5
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    • pp.551-555
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    • 2012
  • Endometriosis is defined as the presence of functioning endometrial tissue outside the uterine cavity, and the standard treatment is extensive surgical excision. Cesarean scar endometriosis is a type of cutaneous endometriosis arising on or near a Cesarean section scar. A 44-year-old woman presented with a $9{\times}6$ cm sized dark-brown, stony-hard, irregular, lower abdominal mass of four years duration. The patient had a history of two Cesarean deliveries, 14 and 16 years ago. Suspecting endometriosis, we excised the tumor conservatively rather than extensively to prevent incisional hernia considering the benign nature of the tumor and the low possibility of recurrence because the patient's age was near menopause, along with simultaneous bilateral salpingo-oophorectomy that was performed in this case. On reconstruction, mini-abdominoplasty was adopted to avoid possible wound complications and cosmetic deformities. The patient was satisfied with the cosmetic results, and neither recurrence nor functional problems occurred during the 1-year follow-up period. Plastic surgeons should keep in mind the possibility of cutaneous endometriosis in an abdominal mass of a female of reproductive age with a previous history of pelvic or intra-abdominal surgery. An optimal result from oncological, functional, and cosmetic standpoints can be achieved with conservative excision followed by mini-abdominoplasty of extensive Cesarean scar endometriosis.

Effect of Prostaglandins on in vitro Oocyte Final Maturation (GVBD) and Ovulation in the Longchin Goby Chasmichthys dolichognathus (점망둑(Chasmichthys dolichognathus)의 최종성숙(GVBD)과 배란 유도에 미치는 Prostaglandins의 영향)

  • Kim, Hyo Eun;Baek, Hea Ja
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.1
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    • pp.41-47
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    • 2017
  • Perhaps the most common type of reproductive dysfunction in captive fish is failure of females to undergo final oocyte maturation and thus to ovulate and spawn. The success of aquaculture could therefore be improved by developing techniques to enhance natural spawning, artificial maturation, and/or to induce ovulation in farmed fish. This study aimed to investigate the effects of prostaglandin $E_2$ ($PGE_2$) and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) on in vitro oocyte maturation (germinal vesicle breakdown, GVBD) and ovulation in the marine fish Chasmichthys dolichognathus. Post-vitellogenic follicles (0.80-0.94 mm diameter oocytes) were incubated with $PGE_2$ or $PGF_{2{\alpha}}$ at concentrations of 5, 50, or 500 ng/mL for 24 hours. A significant increase in GVBD was seen in 0.84 mm and 0.94 mm oocytes incubated with 50 ng/mL $PGE_2$ compared with the control. There was no significant increase in GVBD in any of the other experimental conditions (5 or 500 ng/mL $PGE_2$ or 5, 50, or 500 ng/mL $PGF_{2{\alpha}}$). Neither of the prostaglandins induced ovulation at the concentrations tested.These results suggest that GVBD was induced by incubation with 50 ng/mL $PGE_2$.