No abstract available.
Animals ; Metaphase* ; Mice* ; Oocytes*

The task for chromosome karyotyping and diagnosis is requiring repetitive, time consuming job and high cost even it is done by well-experienced cytogenetists. Therefore an web based chromosome karyotyping instruction system has been established to be able to analyze chromosomes and obtain necessary advises from the database instead of human experts and the database is including 2 divisions with database and agent.For the first of all, database model was constructed with relational database consisting of Patient_DB, image_DB, Disease_DB and Manage_DB. As the second procedure, knowledge base by IF THEN production rule was implemented to a knowledge domain with normal and abnormal chromosomes. For the last, independent agent with the inference by knowledge base could enter the inference data into the database.Experimental data were composed of normal chromosomes of 2,736 patients' cases and abnormal chromosomes of 259 patients' cases that have been obtained from GTG-banding metaphase peripheral blood and amniotic fluid samples.The completed system provides variously morphological information by analysis of normal or abnormal chromosomes and it also makes users enable to control and search the information in a short period with learning of high amount of knowledge.
Amniotic Fluid ; Diagnosis ; Female ; Humans ; Karyotyping* ; Knowledge Bases ; Learning ; Metaphase

Chromosome analysis has been widely used in clinics including prenatal diagnosis. To obtain high-quality metaphase chromosomes, researchers have attempted to modify the methods for chromosome culture, preparation and analysis. Some large research centers also tried to establish standards for quality control. In this paper, modification of methods for the preparation of chromosomes in the last decade is reviewed.
Cells, Cultured ; Chromosomes, Human ; Cytogenetic Analysis ; Humans ; Karyotyping ; Metaphase

Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30-32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1,2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.
Adult ; Cryopreservation ; Female ; Humans ; Metaphase ; Oocytes ; RNA-Seq ; Vitrification

BACKGROUND: Hypersensitivity to both cell-killing and chromosome-damaging effects of ionizing radiation is a consistent feature of cells from individuals with ataxia-telangiectasia (AT). This radiobiological behavior of AT cells is a component of genetic instability and may contribute to cancer risk. Also, heterozygotes for AT-mutated (ATM) genes have no clinical expressions of AT, but may become cancer prone with a moderate increase in in-vitro radiosensitivity. METHODS: We performed a chromosomal analysis on lymphocytes from 3 AT patients, 5 obligate AT carriers (siblings and parents of the patients), and 5 normal controls. RESULTS: Increases in chromosomal breakages after irradiation with 1 gray/min in cells from AT patients ranged from 0.65 to 0.83 rearrangements per metaphase, while in the carriers and controls the levels of breakage were between 0 and 0.15 per metaphase cells (P<0.05). CONCLUSIONS: These results are consistent with previously reported chromosomal radiosensitivity in AT patients. However, carriers do not show moderate radiosensitivity due to various technical factors such as the dose or distance of radiation. Although this research has some limitations due to the small numbers of patients, carriers and controls, this method may be an easy and useful diagnostic tool for AT patients in Korea. (J Korean Neurol Assoc 19(5):509~513, 2001)
Ataxia Telangiectasia* ; Ataxia* ; Chromosome Breakage ; Heterozygote ; Humans ; Hypersensitivity ; Korea ; Lymphocytes* ; Metaphase ; Parents ; Radiation Tolerance ; Radiation, Ionizing

This study was conducted to investigate the effects of Tissue Culture Medium 199 (TCM) and Dulecco's Modified Eagle Medium (DMEM) on the blastulation and grade of human oocytes on Vero cells in vitro. A cohort of 79 and 93 oocytes in metaphase II stage were used in TCM 199 and DMEM respectively. No differences were found in the nurser of oocytes showing two-pronuclei between TCM (82.3%) and DMEM (86.0%). The number of fertilized oocytes reaching the blastocyst was not significant in TCM (60.0%) and DMEM (63.1%).4 total of 89 blastocysts were categorized into the four grades (BG1, BG2, BG3 and early) depending on their morphology. The number of embryos achieving the blastocyst grade 1 (BG1) was significantly higher (p<0.05) in DMEM (50.8%) than TCM (15.0%). It is concluded that cultured oocytes in DMEM with glutamine on Vero cells should be significantly increased BG1.
Blastocyst ; Cohort Studies ; Eagles ; Embryonic Structures* ; Glutamine* ; Humans* ; Metaphase ; Oocytes ; Vero Cells*

Our purpose is to describe a successful twin pregnancy and delivery after intracytoplasmic sperm injection (ICSI) followed by calcium ionophore with spermatozoa from a globozoospermic man. On the second attempt of ICSI, all of eight metaphase II oocytes were fertilized with treatment with calcium ionophore. Day 3 transfer of six normally developing embryos resulted in an ongoing twin pregnancy, and two preterm healthy babies were born in the 33th week of gestation. To the best of our knowledge, this is the first report of pregnancy and delivery after ICSI followed by calcium ionophore with spermatozoa from a globozoospermic man in Korea.
Calcium* ; Embryonic Structures ; Humans ; Metaphase ; Oocytes ; Pregnancy ; Pregnancy, Twin* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa*

OBJECTIVE: To investigate the effects of female age on in vitro maturation and fertilization of immature oocytes from controlled ovarian hyperstimulation (COH) in human IVF-ET program. METHOD: A total of 96 immature oocytes (GV & metaphase I) obtained from 40 cycles of IVF-ET (29 patients). The mean age of female patients was 31.8+/-3.1 years. Ovulation was triggered by urinary or recombinant hCG. Immature oocytes were cultured with YS medium containing 30% of patients' human follicular fluids, LH (1 IU/mL), FSH (1 IU/mL) and EGF (10 ng/mL), and then matured oocytes were fertilized by ICSI. In vitro maturation and fertilization of immature oocytes were analyzed according to age of female (< 34 or > or = 34 years). RESULTS: The maturation rate was similar between two groups (68% vs 64%). The fertilization rate of in?vitro-matured oocytes was higher in patients < 34 years old, but there was no statistical significance (64% vs 50%, p=0.347). The fertilization rate of in-vitro-matured oocytes was significantly lower compared with those of in-vivo-matured oocytes in both age groups (64% vs 79%, p=0.035, 50% vs 86%, p=0.007). CONCLUSION: In older female group, fertilization rate of in-vitro-matured oocytes seems to be decreased. Further investigations should be warranted to increase fertilization potential of in-vitro-matured oocytes.
Adult ; Epidermal Growth Factor ; Female ; Fertilization* ; Follicular Fluid ; Humans* ; Metaphase ; Oocytes* ; Ovulation ; Sperm Injections, Intracytoplasmic

Sister chromatid exchanges(SCEs) and cell cycle kinetics were proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity in short-term cultures of phytohemagglutinin(PHA)-stimulated human lymphocytes. Therefore, this study was performed to investigate the relation between the cytotoxic effects and sister chromatid exchanges. The results are summarized as follows: 1) The frequency of SCEs per cell are 13.1+/-2.8 in the lower concentration of 6.25x10(-9) M and 75.8+/-8.2 in the highest concentration of 1.00+/-10(-7) M. Mitotic index is decreased in the higher concentration of mitomycin C. The result indicates that mitomycin C led to a dose dependent increase in SCE frequency, but decrease in mitotic index. 2) Chromosomal analysis was performed on metaphase cells that have divided one, two, and three or more times for cell cycle kinetics by fluorescence-plus-Giemsa(FPG) technique. According to the increased but the cells of third division are greatly decreased. 3) The frequency of SCEs per chromosome by chromocomal group are decreased gradually from A group to G group. But relationships between specific chromosomal group and SCEs frequency are not found.
Cell Cycle ; Chromatids ; Humans ; Humans* ; Kinetics ; Lymphocytes ; Metaphase ; Mitomycin* ; Mitotic Index ; Siblings* ; Sister Chromatid Exchange*

OBJECTIVE: The purpose of this study was to evaluate the efficacy and effect of various cryopreservation method on the survival and the cytoskeletal stability of metaphase II mouse oocyte. METHODS: Mouse ovulated oocytes were collected and cryopreserved by a modified slow-freezing method with 1.5 M 1,2-propanediol (PrOH)+0.1 M sucrose or by vitrification using cryo loop and EM grid with 40% ethylene glycol+0.6 M sucrose. Four hours after thawing, intact oocytes were fixed and stained with fluorescein isothiocyanate (FITC)-conjugated monoclonal anti-beta-tubulin antibody to visualize spindle and propidium iodide (PI) to visualize chromosome. Spindle morphology was classified as follows: normal (barrel-shaped), slightly and absolute abnormal (multipolar or absent). RESULTS: Survival rate of the frozen-thawed oocytes in vitrification group was significantly higher than that of slow-freezing group (62.7% vs. 24.4%, p<0.01). Vitrification with cryo loop showed significantly higher survival rate than that with EM grid (67.7% vs. 53.5%, p<0.05). On the other hand, proportion of normal spindle and chromosome configurations of the frozen-thawed oocytes between two vitrification group was not significantly different. CONCLUSION: For mouse ovulated oocytes, vitrification with cryo loop may be a preferable procedure compared to slow-freezing method. Further study should be needed to investigate developmental competency of frozen-thawed mouse oocytes.
Animals ; Cryopreservation* ; Fluorescein ; Hand ; Metaphase ; Mice* ; Oocytes* ; Propidium ; Propylene Glycol ; Sucrose ; Survival Rate ; Vitrification