RESUMO
PURPOSE: The present study was undertaken to evaluate the effects of morphokinetic abnormalities of human spermatozoa on chromatin packing and DNA integrity and possible beneficial effects of sperm selection in ICSI. METHODS: Semen samples from 1002 patients were analysed for morphology and motility using CASA. Protamine status and DNA fragmentation were analysed by chromomycin A3 staining and sperm chromatin dispersion assay respectively. RESULTS: Sperms with elongated, thin, round, pyri, amorphous, micro and macro forms were significantly higher in teratozoospermic and oligoasthenoteratozoospermic groups. Significant difference in chromatin packing and DNA fragmentation index was observed in these abnormal groups compared with normal. Similarly significant correlation was also seen between abnormal motility parameters and DNA fragmentation index in asthenozoospermic group compared with normal. CONCLUSIONS: Specific abnormal morphological forms have higher incidence of chromatin packing abnormalities and DNA fragmentation. Using these sperms in ICSI might have an impact on fertilization, embryo development and abortion rates. These can be selectively avoided during ICSI procedure to improve ART outcome.
Assuntos
Cromatina , Fragmentação do DNA , Infertilidade Masculina , Espermatozoides , Adulto , Antígenos de Neoplasias/metabolismo , Cromatina/genética , Cromatina/ultraestrutura , Cromomicina A3 , Desenvolvimento Embrionário , Feminino , Humanos , Infertilidade Masculina/genética , Infertilidade Masculina/fisiopatologia , Masculino , Pessoa de Meia-Idade , Gravidez , Protaminas/metabolismo , Injeções de Esperma Intracitoplásmicas/métodos , Espermatozoides/anormalidades , Espermatozoides/citologiaRESUMO
PURPOSE: The present study was undertaken to evaluate and compare the post thaw survival, implantation and pregnancy rates of vitrified human early cavitating blastocysts with deflated expanded blastocysts. MATERIAL AND METHODS: Supernumerary blastocysts were vitrified in 30% ethylene glycol-dimethyl sulphoxide based solution using cryoloop. Fully expanded blastocysts were deflated by gentle aspiration of the blastocoelic fluid using a micromanipulator until the cavity collapses prior to vitrification. RESULTS: Of the 576 vitrified blastocysts, 545 (94.61%) survived thawing in the early cavitating blastocyst group which was significantly higher than deflated expanded blastocyst group, in which only 370 survived thawing out of 459 (80.62%). However, no significant difference was observed in implantation and pregnancy rates between early cavitating and deflated expanded blastocyst groups. CONCLUSIONS: Early cavitating blastocyst would be the ideal stage for cryopreservation of human blastocysts as it has higher survival rate and avoids additional invasive procedures like deflation of the blastocoele.
Assuntos
Blastocisto/fisiologia , Criopreservação/métodos , Implantação do Embrião , Taxa de Gravidez , Blastocisto/citologia , Blastocisto/efeitos dos fármacos , Técnicas de Cultura , Dimetil Sulfóxido/farmacologia , Etilenoglicol/farmacologia , Feminino , Humanos , Gravidez , Análise de SobrevidaRESUMO
A binary system for gene activation and site specific integration based on conditional recombination of transfected sequences mediated by FLP recombinase from yeast was implemented in mammalian cells. In several cell lines, FLP rapidly and precisely recombined copies of its specific target sequences to activate an otherwise silent beta-galactosidase reporter gene. Clones of marked cells were generated by excisional recombination within a chromosomally integrated copy of the silent reporters. These clones exhibited intense blue colour with X-Gal staining solution.
