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: Chromosomal aberration observed only in a few metaphases may cause the cytogeneticists to have difficulties in making a decision whether it is due to in vivo mosaicism/multiple clones or due to in vitro artifact. This is especially important when the chromosome of concern has been associated with a classical chromosome syndrome, malignancy or its evolution. Therefore, we aimed to establish a range for random chromosomal aberrations among cells from PHA-stimulated blood(PB) and bone marrow(BM) cultures. METHODS: Among the cells from 449 PB and 472 BM specimens referred for chromosome studies from 1997 to 1998, we analyzed the frequency of random aneuploidy, structural abnormalities, and breaks/gaps. RESULTS: The number of cells analyzed was 5,904/4,488(1997/1998) in PB and 4,211/4,124(1997/1998) in BM. The frequency of metaphases with random chromosomal aberrations of BM(32.10%) was much higher than that of PB(5.90%). The most frequent aberration was chromsome loss. Autosome losses were inversely correlated with autosome size(correlation coefficient = -0.83 and -0.72, p<0.01), smaller chromosomes being lost more frequently while autosome breaks/gaps were correlated with autosome size(correlation coefficient = 0.69 and 0.85, p<0.01), in PB and BM. Comparing the data from 1998 to the data from 1997, the frequency of chromosome losses(<0.5% in PB, <2.25% in BM), gains(<0.1% in PB and BM), breaks/gaps(<0.1% in PB, <0.25% in BM), and structural aberrations( Aneuploidy ; Artifacts ; Bone Marrow* ; Chromosome Aberrations* ; Chromosomes, Human, 4-5 ; Clone Cells ; Metaphase ; Quality Control

BACKGROUND: The KCl hypotonic treatment is important for swelling the cells and adequate spreading of chromosomes on the slide. Cytogenetic laboratory usually use 0.075M KCl solution. Sometimes, it is difficult to obtain enough and good quality of metaphase cells, because of inadequate hypotonic treatment. The purpose of this study was to evaluate the mitotic index and band resolution according to the different KCl concentration. METHODS: The group I included blood specimens obtained from 14 newborns (median age 1 day, range 1-8 days) and 4 cord blood. The group II included 16 persons whose median age was 28 years (1-37 years). The blood was cultured in RPMI 1640 medium with fetal calf serum and phytohemagglutinin for 72 hours. Mitosis was arrested by adding colcemid (100 ng/mL). The hypotonic treatment was done by adding different KCl concentration such as 0.075M, 0.068M and 0.057M for 30 minutes at 37 degrees C. The mitotic index was calculated as the number of metaphase cells per total 1,000 cells. The band resolution was evaluated by 2 persons independently. RESULTS: For group I, the mitotic index was not different according to the KCl concentration; 0.075M, 18.8 (5.5~31.5); 0.068M, 22.3 (11~32.5); 0.057M, 20.5 (2.5~29), (P=0.137). The proportion of cells with 400 or more band resolution was significantly higher in specimens treated with 0.068M KCl than those treated with 0.075M KCl; 0.075M, 67.8% (56~92.5); 0.068M, 73.6% (46.1~84.6); 0.057M, 71.6% (63~89.2), (P=0.027). For group II, the results were similar to those of group I. The mitotic index was as follows; 0.075M, 22.3 (5~28); 0.068M, 26 (4~34.5); 0.057M, 21.5 (2.5~36.5), (P=0.568). The proportion of cells with 400 or more band resolution was as follows; 0.075M, 66.6% (42.8~83.3); 0.068M, 69.7% (54.3~87.5); 0.057M, 68.2% (50~78.6) (P=0.04). CONCLUSIONS: For 0.068M or 0.057M KCl treatment, band resolution was improved, while the mitotic index was similar to that of 0.075M KCl. We suggest use of 0.068M or 0.057M KCl hypotonic treatment in addition to 0.075M KCl for chromosome preparation of peripheral blood.
Cytogenetics ; Demecolcine ; Fetal Blood ; Humans ; Infant, Newborn ; Metaphase ; Mitosis ; Mitotic Index*

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

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*

BACKGROUND: One of major challenge of cytogenetics is to obtain qualitative metaphases to achieve a meaningful analysis. Methotrexate (MTX) synchronization and the Giant Cell Tumor-Conditioned Medium (GCT-CM) have been used to improve the metaphase preparation from hematopoietic malignancies. The purpose of this study is to determine Mitotic Index (MI) of bone marrow samples under several culture conditions that may improve the quality of chromosome preparation. METHODS: Sixty nine bone marrow samples were cultured into 3 groups by traditional methods. The first group was tested for the effect of cell concentration on MI. The second, for the effect of MTX concentration on MI. The third group was classified into 4 subgroups as follows: 1) MTX only in 24 hour culture 2) MTX and GCT-CM in 24 hour culture 3) 48 hour culture without MTX 4) 48 hour culture and GCT-CM. MI was calculated as the ratio of metaphase to interphase cells in 1000 cells. Quality of metaphase was evaluated by classified the metaphase cell into 3 types. RESULTS: The first and second groups revealed no relationship between cell concentration, amount of MTX and MI, respectively. The third group showed significant differences among four subgroups. The MI increased in subgroups using GCT-CM and in the 48 hour culture, with greatest increase in group using 48 hour culture and GCT-CM simultaneously. CONCLUSIONS: The use of GCT-CM medium and prolonged culture improved the quality of metaphase cells and MI. It is therefore beneficial to use these conditions in cytogenetic studies on bone marrow in hematologic disease.
Bone Marrow* ; Cytogenetics ; Giant Cells* ; Hematologic Diseases ; Hematologic Neoplasms ; Interphase ; Metaphase ; Methotrexate ; Mitotic Index*