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Effect of Fertilization Promoting Peptide on Kinematic Parameters, Capacitaion and Acrosome Reaction in Human Spermatozoa.

Moon Kyoo KIM ; Hee Gyoo KANG ; Myo Kyung KIM ; Dong Hoon KIM ; Sung Won HAN ; Do Hyun CHOI ; Ho Hoon LEE

Korean Journal of Fertility and Sterility.2000;27(2):201-208.

OBJECTIVE: This study has been carried out to evaluation the effect of fertilization promoting peptide (FPP) on the kinematic parameters, capacitation and acrosome reaction of the frozen-thawed human spermatozoa. METHODS: After FPP treatment, we examined kinematic parameters, capacitation and acrosome reaction, using the methods of computer-aided sperm analysis (CASA) and chlortetracycline (CTC) fluorescence analysis. RESULTS: We have obtained the evidence that FPP can promote the capacitation and inhibit the spontaneous acrosome reaction of frozen-thawed human spermatozoa in vitro. Fpp (25~100 nM) induced a significant increase in the proportion of B-pattern capacitated spermatozoa, and a significant decrease in the proportion of F-pattern uncapacitated ones without significant stimulation of acrosomal exocytosis. In the kinematic parameters treatment, FPP treated groups maintained higher LIN, BCF and STR than those of control. The VAP, VSL, VCL and ALH were not different. Therefore it is suggested that FPP in human seminal plasma may play a positive role in promoting human sperm function.
Acrosome Reaction* ; Acrosome* ; Chlortetracycline ; Exocytosis ; Fertilization* ; Fluorescence ; Humans* ; Male ; Semen ; Spermatozoa*

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Effects of the Stepwise Exposure Treatments Before Freezing on the Survival Capacity of the Frozen-Thawed Mouse Mature Oocytes by Vitrification or Ultra-Rapid Freezing.

Sang Woo KIM ; Young Ah LEE ; Man Soo YOON ; Kyu Sup LEE ; Jae Ik LEE ; Mi Kyung KIM

Korean Journal of Fertility and Sterility.2000;27(2):191-200.

OBJECTIVE: This study was carried out to compare the effects of the stepwise exposure treatments on the morphological normality, fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing and to use as a fundamental data for the cryopreservation of human oocytes. MATERIALS AND METHODS: The morphological normality and fertilization rates of the vitrified and ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were observed. After choosing the 3step exposure treatment groups, we observed the morphological normality and fertilization, blastocyst formation rate vitrified and ultra-rapid frozen mouse mature oocytes. RESULTS: The morphological normality and fertilization rates of the vitrified mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 75%, 85%, 88% and 58%, 61%, 54% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rates of the control was 92% and 65%. There were no significant differences in fertilization rate among control and treatments (p>0.05). The morphological normality and fertilization rates of the ultra-rapid frozen mouse mature oocytes after three-stepwise exposure treatments (1step, 3step and 5step) were 83%, 83%, 84% and 75%, 63%, 56% respectively. There were no significant differences among treatments (p>0.05). The morphological normality and fertilization rate of the control was 95% and 67%. There were no significant differences among control and treatments (p>0.05). The morphological normality and fertilization rate of the vitrified or ultra-rapid frozen mouse mature oocytes after 3step exposure treatment were 69% and 75%, respectively. The blastocyst formation rate was 60% and 57%. The results did not differ significantly between vitrification and ultra-rapid freezing (p>0.05). CONCLUSION: As known in the above results, there were no significant differences in the fertilization and blastocyst formation rate of the frozen-thawed mouse mature oocytes by vitrification or ultra-rapid freezing among the control and treatments. It is suggested that vitrification and ultra-rapid freezing method were effective for the cryopreservation of mouse mature oocytes.
Animals ; Blastocyst ; Cryopreservation ; Fertilization ; Freezing* ; Humans ; Mice* ; Oocytes* ; Vitrification*

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Regulation of LIF Gene Expression by Interleukin-1 in the Mouse Peri-implantation Embryos and Uterine Endometiral Cells.

Joung Woul KIM ; Hye Young YANG ; Hyoung Eun RYU ; Ji Youn LEE ; Moon Kyoo KIM ; Jung Bok LEE ; Hyun Soo YOON

Korean Journal of Fertility and Sterility.2000;27(2):183-190.

No abstract available.
Animals ; Embryonic Structures* ; Gene Expression* ; Interleukin-1* ; Mice*

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The Analysis of LHbeta -subunit Variants in Infertile Patients with Premature Ovarian Failure (POF) in Korea.

Eu Gene LEE ; Nam Keun KIM ; Sook Hwan LEE ; Jung Jae KO ; Kwang Yul CHA ; Hye Sun JEON ; Kyung Ah LEE

Korean Journal of Fertility and Sterility.2000;27(2):179-182.

This study was performed to determine whether the LHbeta -subunit gene missense mutation is present in Korean infertile patients with 46,XX POF women. The variants of LHbeta exon 2 (Trp 8Arg; TGG to CGG and Ile15Thr; ATC ti ACC) were studied in forty-four 46.XX idiopathic POF and 54 nonpregnant women. The LHbeta exon 2 variants were more frequent in POF patients (20.5%) than nonpregnant( 16.7%) women (p>0.05). POF patients with the variant was slightly higher than nonpregnant women with the variant.
Exons ; Female ; Humans ; Korea* ; Mutation, Missense ; Primary Ovarian Insufficiency*

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Molecular Variants of the LHbeta-subunit in Patients with Polycystic Ovary Syndrome (PCOS) in Korean Women.

Eu Gene LEE ; Tae Jong SOHN ; Nam Keun KIM ; Sook Hwan LEE ; Jung Jae KO ; Kwang Yul CHA ; Hyung Min CHUNG

Korean Journal of Fertility and Sterility.2000;27(2):173-178.

No abstract available.
Female ; Humans ; Polycystic Ovary Syndrome*

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Subsequent Embryo Transfers (SET) on Day 2 and 5: It's Safety and Effectiveness.

Taek Hoo LEE ; Sang Sik CHUN ; Kee Sang PARK ; Hai Bum SONG

Korean Journal of Fertility and Sterility.2000;27(2):165-172.

OBJECTIVE: In vitro fertilization (IVF) and a prolonging the time of culture may be helpful in establishing a viable pregnancy through a selection effect. Some embryos do not develop beyond the 4-cell stage and some may not develop to the blastocyst stage. We have evaluated the safety of SET and the outcomes of pregnancy. METHODS: Sperms were treated with Ham's F-10 supplemented with 10% human follicular fluid (hFF). Oocytes or fertilized oocytes were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% or 20% hFF respectively. Up to five oocytes were inseminated with approximately 200,000 sperm cells/2 ml in each well. Fertilization was examined in the following morning and fertilized oocytes were co-cultured until embryo transfer. Vero cells for co-culture were prepared in Tissue Culture Medium - 199 (TCM-199) with 10% fetal bovine serum. At the two to four cell and blastocyst on day 2 and day 5, embryo and blastocyst grading were evaluated. Pregnancy rate was determined after transfer of human embryos at the two to four cell stage on day 2 (Group I) or subsequent transfer of embryos on day 2 and at the blastocyst stage on day 5 (Group II). For statistically significant when rho value was less than 0.05. RESULTS: No differences was found in the fertilization between Group I(81.0%, 98/121) and Group II(81.8%, 180/220). In case of cleavage rate, no difference was found in Group I(95.9%, 94/98) and Group II(66.7%, 12/18) than in Group (26.3%, 5/19). CONCLUSION: The results of this study showed that SET is safe and effective, and significantly increases the pregnancy rate.
Blastocyst ; Coculture Techniques ; Eagles ; Embryo Transfer* ; Embryonic Structures* ; Female ; Fertilization ; Fertilization in Vitro ; Follicular Fluid ; Humans ; Oocytes ; Pregnancy ; Pregnancy Rate ; Spermatozoa ; Vero Cells

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Effect of Hydrosalpingeal Fluid on the Implantation in-vitro in a Murine Model.

Jin Hyun JUN ; Chun Kyu LIM ; Soo Kyung KIM ; Mi Kyoung KOONG ; Inn Soo KANG

Korean Journal of Fertility and Sterility.2000;27(2):159-164.

No abstract available.

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The Effect of Polycystic Ovarian Follicular Fluid on Sperm Motility in Human in vitro Fertilization.

Yeon Hee KIM ; Sang Hoon LEE ; Min HUR

Korean Journal of Fertility and Sterility.2000;27(2):151-158.

OBJECTIVE: The purpose of this study was to evaluate the effect of polycystic ovarian follicular fluid on sperm motility in human in vitro fertilization (IVF). METHODS: From May, 1998 to July, 1999, 55 patients who complained of infertility were involved in this study. We obtained ovarian follicular fluids form the patients by ultrasono-guided aspiration. Subjects were divided into two groups. 20 patients who had polycystic ovarian disease were belong to study group, and 25 patients who had normal ovarian follicular fluid were belong to control group. The follicular fluid dilution was done with Ham's fluid as 10%, 20%, 50%, 100%. The sperm motility was analyzed by CASA at 6hr and 12hr after incubation in follicular fluids. RESULTS: The levels of average path velocity (VAP) in all concentration fluid didn't show significant difference between study and control group. The other parameters including curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), and linerity (LIN) were didn't show any significant difference between both groups. CONCLUSION: PCOD fluid had seemed to have an adverse effect on the sperm biological function. But, this study showed that PCOD fluid had no different effect on sperm motility with normal follicular fluid.
Female ; Fertilization in Vitro* ; Follicular Fluid* ; Head ; Humans* ; Infertility ; Ovarian Diseases ; Sperm Motility* ; Spermatozoa*

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Xenografting of the Human Vitrified Ovarian Tissues into the Immune Deficient Animal.

Se Jin YOON ; Sook Hyun LEE ; Chang Sook SHIN ; Kyung Ah LEE ; Yong Seon CHO ; Tae Ki YOON ; Kwang Yul CHA

Korean Journal of Fertility and Sterility.2000;27(2):145-150.

OBJECTIVE: The present study was conducted to evaluate the viability of germ cells form the adult and fetal ovarian tissues after vitrification followed by xenografting. METHOD: The human adult ovarian tissues were obtained from 33 years old patients, and the fetal ovarian tissues were obtained from 22 weeks and 25 weeks in gestation. ovarian tissues were cryopreserved by vitrification with 5.5 M ethylene glycol (EG 5.5) and 1.0 M sucrose as cryoprotectants. Adult and fetal ovarian tissues were pre-equilivrated with EG 5.5 at room temperature for 10 and 5 minutes, respectively and plunged into liquid nitrogen immediately. Frozen-thawed tissues were xenografted into NOD-SCID mice to evaluated the viability and capacity for further growth of the primordial follicles. Grafts were recovered from the recipients 4 weeks after transplantation and histological analysis was accomplished. RESULT AND CONCLUSION: Grafts recovered 4 weeks after transplantation contained less number of oocytes and primordial follicles compared to that of the fresh tissues. Survived follicles were mainly primordial and intermediary with larger diameter and more granulosa cells. It is confirmed that 1) the ovarian tissues were healthy and the germ cells were survived after vitrification, and 2) the survived fetal primordial follicles after vitrification resumed the growth in the xenofrafts.
Adult ; Animals* ; Ethylene Glycol ; Female ; Germ Cells ; Granulosa Cells ; Heterografts* ; Humans* ; Mice ; Nitrogen ; Oocytes ; Pregnancy ; Sucrose ; Transplantation, Heterologous* ; Transplants ; Vitrification

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Molecular Identification and Distribution of Aquaporins in Human and Rat Testes.

Nam Cheol PARK ; Young Soo PARK ; Jin Sup JUNG

Korean Journal of Fertility and Sterility.2000;27(2):133-144.

OBJECTIVE: Several water channels (aquapoins; AQP) that belong to the MIP (major intrinsic protein) family have identified. In the selected tissues including red blood cells or renal tubules, water movements are abundant and/or physiologically important. Unexpectedly, a high water permeability of human and ram sperm has been reported. Recent studies showed that AQP7 and AQP8 are present in testes so that the high water permeability of human sperm suggested to be mediated by AQPs. METHOD: To identify the identity of aquaporins expressed in testes, RT-PCR was performed using degenerative primers, which were designed to correspond to highly conserved sequences surrounding the Asn-Pro-Ala (NPA) motifs in the aquaporins. New expressed AQP series were reconfirmed by immunohistochemical study using rabbit polyclonal antibodies. RESULTS: DNA sequencing of PCR products revealed that AQP2 and AQP3 mRNA as well as AQP7 and AQP8 are expressed in human and rat testes, AQP2 are expressed in spermatozoa, interstitial cells and myofibroblasts and AQP3 are expressed in myofibroblasts of semineferous tubules on immunocytochemical stain. CONCLUSION: These results indicate that multiple aquaporins are expressed in testes, and that they may have important roles in the spermatogenesis and the germ cell function of testis.
Animals ; Antibodies ; Aquaporins* ; Conserved Sequence ; Erythrocytes ; Germ Cells ; Humans* ; Myofibroblasts ; Permeability ; Polymerase Chain Reaction ; Rats* ; RNA, Messenger ; Sequence Analysis, DNA ; Spermatogenesis ; Spermatozoa ; Testis* ; Water Movements

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