Blastomeres; Serum; Pathological Conditions, Anatomical; Chorionic Gonadotropin The group 1 of 75 cases of post molar evacuation and the group 2 of 292 cases of gestational trophoblastic tumors (GTT) were followed up until beta-hCG level serum, determined by IMX, reaches the threshold of < 5 IU/l, there was not any complication after 2 years. There were 7.5% recurrents cases in GTT group, in some cases, beta-hCG reaches 0 UI/l. In the recurrent group, the rate of resistance to the treatment got to 40%, and the rate of resistance to single chemotherapy by MTX got to 30%. 50% of recurrent patients have had uterine histological exams with 36% choriocarcinoma, 4.5% invasive molar
Blastomeres ; Serum ; Pathological Conditions, Anatomical ; Chorionic Gonadotropin

Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Zl, DXZI, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.
Blastomeres ; Chromosome Aberrations ; DNA Probes ; Fluorescence* ; Germ Cells ; Humans ; In Situ Hybridization* ; Interphase ; Lymphocytes ; Metaphase ; Spermatozoa

OBJECTIVE: To compare the mouse oocyte vitrification outcomes of the CryoLogic vitrification method (CVM) and the conventional open method using a Cryotop. Two CVM methods (original CVM and modified CVM) were tested. METHODS: Mature oocytes obtained from female BDF-1 mice were vitrified by two-step exposure to equilibrium and vitrification solutions. Three vitrification protocols were tested on three groups: the CVM-kit, modified CVM, and Cryotop groups. After exposure to the two solutions, the oocytes were vitrified. After warming, the oocytes were fertilized in vitro, and the embryo development was assessed. Blastomeres positive for caspase were counted using an in situ assay kit. The spindle morphology and chromosome configurations of warmed vitrified oocytes were also assessed. RESULTS: The modified CVM and Cryotop groups showed similar developmental capacities, and similar proportions of cells with intact spindles and chromosome configurations. The modified CVM protocol was superior to the original CVM protocol for developmental competence and intact spindle preservation. However, the CVM group showed a relatively higher number of apoptotic cells in blastocysts. CONCLUSION: Closed vitrification using the modified CVM protocol may be used as an alternative to the conventional open method, but strategies to decrease apoptosis in the blastomere need to be investigated.
Animals ; Apoptosis ; Blastocyst ; Blastomeres ; Embryonic Development ; Female ; Humans ; Mental Competency* ; Methods ; Mice* ; Oocytes* ; Pregnancy ; Vitrification*

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'toysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.
Blastomeres ; Chorionic Villi ; Diagnosis ; DNA ; Dystrophin* ; Embryonic Structures ; Exons ; Genome ; Oocytes ; Polar Bodies ; Spermatozoa

OBJECTIVE: Human infertility clinics have been faced the demand for improving clinical results. The purpose of this study was to evaluate the effect of microsurgical removal of damaged blastomeres (DB) in frozen-thawed embryos on the clinical outcomes. METHODS: From January 2003 to May 2004, out of 258 thawing ET cycles were divided into three groups: Group-1 (n=46): Intact cleavaged embryos after thawing. Remained cycles with embryos containing DB were randomly divided into two groups. Group-2 (n=102): Drilling zona pellucida (ZP) of frozen-thawed embryos by acidified Tyrode's solution. Group-3 (n=110): Drilling ZP and removal of DB. Embryos after microsurgical manipulation were transferred into the uterus of patients. RESULTS: Clinical profiles and the mean number of transferred embryos among three groups were not different. Pregnancy and implantation rates were similar in three groups. It were 30.4% and 9.3% in Group-1, 29.4% and 7.8% in Group-2, and 26.4% and 7.6% in group-3, respectively. Miscarriage rate in Group-3 (37.9%) was slightly higher than those in Group-1 and Group-2 (14.3% and 23.3%), but it was not statistically significant. CONCLUSION: Intact cleaving embryos after DB removal showed higher potent of pregnancy and implantation. We could not find any improvement of clinical outcome by removal of DB in frozen-thawed embryos.
Abortion, Spontaneous ; Blastomeres* ; Embryo Transfer* ; Embryonic Structures* ; Female ; Humans ; Infertility ; Pregnancy ; Pregnancy Rate ; Uterus ; Zona Pellucida

OBJECTIVE: To investigate the beneficial effect of fragment removal on the subsequent cell division and clinical outcome of the fragmented human embryos. METHODS: A prospective study was performed in Hanna Women's Clinic and Mizmedi Hospital. Sixty couples undergoing In vitro fertilization-embryo transfer (IVF-ET) program were participated in the present study. The microsurgical fragment removal was performed in 106 fragmented embryos of 29 patients before the transfer. As a control group, 122 fragmented embryos of 31 patients were transferred without the fragment removal. Effects of fragment removal on morphological changes and clinical outcomes of fragmented embryos were investigated. RESULTS: Mean morphological grade (G2.79) of fragmented embryos was significantly improved after the fragment removal (G1.63, p<0.001). Most of the fragmented embryos did not show a regeneration of fragments after the fragment removal during the subsequent development, and a beneficial effect of fragment removal on the development of the fragment removed embryos was observed. Implantation and pregnancy rates of fragment removed embryos were 12.3% and 31.3%, whereas the rates of control group embryos were 6.6% and 22.5%, respectively. There was no statistical significance in the rates between the two groups because of the low number of trials. CONCLUSION: Microsurgical fragment removal improved the subsequent development as well as the morphological grade of fragmented embryos. The fragment removal may be beneficial for neighboring blastomeres by repairing the intercellular communication and removing the secretion of the potential toxic materials by fragments.
Blastomeres ; Cell Division ; Embryonic Structures ; Family Characteristics ; Humans ; Pregnancy Rate ; Prospective Studies ; Regeneration

PURPOSE: To determine a method to improve the efficacy and accuracy of preimplantation genetic diagnosis (PGD) - polymerase chain reaction (PCR), we compared hot start PCR and conventional multiplex nested PCR. MATERIALS AND METHODS: This study was performed with single lymphocyte isolated from whole blood samples that were obtained from two couples with osteogenesis imperfecta (OI). We proceeded with conventional multiplex nested PCR and hot start PCR in which essential reaction components were physically removed, and we compared the amplification rate, allele dropout rate and nonspecific products. Afterward, we used selective method for PGD. RESULTS: In the two couples, the respective amplification rate were 93.5% and 80.0% using conventional multiplex nested PCR and 95.5% and 92.0% using hot start PCR. The respective mean allele dropout rates for the two couples were 42.0% and 14.0% with conventional multiplex nested PCR and 36.0% and 6.0% with hot start PCR. CONCLUSION: The results demonstrate that the hot start PCR procedure provides higher amplification rates and lower allele dropout rate than the conventional method and that it decreased the nonspecific band in multiplex nested PCR. The hot start method is more efficient for analyzing a single blastomere in clinical PGD.
Alleles ; Blastomeres ; Family Characteristics ; Humans ; Lymphocytes ; Osteogenesis Imperfecta ; Patient Dropouts ; Polymerase Chain Reaction ; Preimplantation Diagnosis ; Prostaglandins D

It is well known that developmental delay or arrest occurs before implantation inmammals, which have undergone in vitro culture. Recently, these phenomenon are being attributedto oxygen free radicals, and successful cell culture are being obtained by lowering theoxygen environment of in vitro culture. This is due to the fact that the oxygen concentrationin the fallopian tube is around 5%, which is lower than the room air 20% concentrationfor in vitro culture.Rutin, which is a free radical scavenger, was added to early embryo mice culture andcompared the free radical level at blastocyst stage with that of different culture conditions,and found that free radical level was markedly decreased. Also, there was increased embryodevelopment with decreasing free radical levels in the experimental group, and there wassignificant increase in the blastulation rate and blastomere count.This study therefore suggests the possibility of improved in in-vitro embryo culture.
Animals ; Blastocyst* ; Blastomeres ; Cell Count* ; Cell Culture Techniques ; Embryonic Structures* ; Fallopian Tubes ; Female ; Free Radicals ; Mice* ; Oxygen ; Rutin*

It is well known that developmental delay or arrest occurs before implantation inmammals, which have undergone in vitro culture. Recently, these phenomenon are being attributedto oxygen free radicals, and successful cell culture are being obtained by lowering theoxygen environment of in vitro culture. This is due to the fact that the oxygen concentrationin the fallopian tube is around 5%, which is lower than the room air 20% concentrationfor in vitro culture.Rutin, which is a free radical scavenger, was added to early embryo mice culture andcompared the free radical level at blastocyst stage with that of different culture conditions,and found that free radical level was markedly decreased. Also, there was increased embryodevelopment with decreasing free radical levels in the experimental group, and there wassignificant increase in the blastulation rate and blastomere count.This study therefore suggests the possibility of improved in in-vitro embryo culture.
Animals ; Blastocyst* ; Blastomeres ; Cell Count* ; Cell Culture Techniques ; Embryonic Structures* ; Fallopian Tubes ; Female ; Free Radicals ; Mice* ; Oxygen ; Rutin*

OBJECTIVE: To determine whether the serum beta-human chorionic gonadotropin (hCG) profile following preimplantation genetic diagnosis (PGD) is lower than that of intracytoplasmic sperm injection (ICSI) cycles. METHODS: A total of 129 PGD cycles and 1,161 age-matched ICSI cycles, which resulted in pregnancy (serum beta-hCG> or =5 mIU/mL) on post-ovulation day (POD) 12 were included. We compared the mean serum beta-hCG levels on POD 12, 14, 21, and 28, doubling time of serum hCG, and created a cut-off value for predicting a singleton pregnancy in each group. RESULTS: The mean serum beta-hCG concentration of the PGD group was significantly lower than that of the control group on POD 12, 14, and 21. The doubling time of serum beta-hCG at each time interval showed no significant difference. The cut-off-value of serum beta-hCG for predicting a single viable pregnancy was 32.5 mIU/mL on POD 12 and 113.5 mIU/mL on POD 14 for the PGD group, which was lower than that for the control group. CONCLUSION: Blastomere biopsy may decrease the beta-hCG-producing activity of the trophoblasts, especially in early pregnancy. Setting a lower cut-off value of serum beta-hCG for predicting pregnancy outcomes in PGD may be needed.
Biopsy ; Blastomeres ; Chorionic Gonadotropin ; Female ; Humans ; Pregnancy ; Pregnancy Outcome ; Preimplantation Diagnosis ; Prostaglandins D ; Sperm Injections, Intracytoplasmic ; Trophoblasts