The aim of this study was to evaluate the efficiency of using meiotic spindle (MS) visibility and relative position to the polar body (PB) as indicators of oocyte maturation in order to optimize intracytoplasmic sperm injection (ICSI) timing. This was a cohort study of patients younger than 40 years with planned ICSI, the timing of which was determined by MS status, compared with those without MS evaluation. The angle between PB and MS and MS visibility were evaluated by optical microscope with polarizing filter. Oocytes with MS evaluation were fertilized according to MS status either 5-6 h after ovum pick-up (OPU) or 7-8 h after OPU. Oocytes without MS evaluation were all fertilized 5-6 h after OPU. For patients over 35 years visualization of MS influenced pregnancy rate (PR): 182 patients with MS visualization had 32% PR (58/182); while 195 patients without MS visualization had 24% PR (47/195). For patients under 35 years, visualization of MS did not influence PR: 140 patients with MS visualization had 41% PR (58/140), while 162 patients without MS visualization had 41% PR (66/162). Visualization of MS therefore appears to be a useful parameter for assessment of oocyte maturity and ICSI timing for patients older than 35.
For group of 281 oocytes obtained from 43 stimulated donors and cryopreserved by vitrification protocol using Cryotop and Kitazato medium we determined important parameters of oocytes collection and vitrification processes which strongly affect the probability that warmed oocytes will produce high-quality embryos for transfer. The probability to obtain high-quality embryos for transfer from vitrified and warmed oocytes was highest when two conditions were fulfilled: 1. oocytes were incubated before vitrification for 7-10 h and 2. stimulated ovaries of donors in one cycle produced a smaller number of oocytes (<7 oocytes from one donor per stimulated cycle). The probable reasons for these observations were: 1. early vitrification (less than 7 h) before final oocyte metaphase II maturation negatively affected the crucial process of post-warm remodelling of spindles and chromosomes, which reduced the fertilization and utilization rates, 2. the evaluated vitrification protocol amplifies negative impact of membrane defects of oocytes of those cohorts containing more than 6 oocytes - freezing places great demands on the integrity and elasticity of the cell membranes. The fact that cryopreservation influences a complex state of oocytes was confirmed by confocal microscopy.
OBJECTIVE: To determine whether varied human spermatozoa, as detected with monoclonal antibodies against acrosomal proteins, have an influence on fertilization, transfer, pregnancy, and implantation rates when intracytoplasmic sperm injection is used. DESIGN: A retrospective study. SETTING: A private IVF center and academic research laboratory. PATIENT(S): One thousand two hundred forty men participating in the intracytoplasmic sperm injection program. INTERVENTION(S): Sperm were divided into seven groups: oligozoospermia, oligoasthenozoospermia, and oligoasthenoteratozoospermia and fresh and frozen-thawed epididymal and fresh and frozen-thawed testicular sperm. Fertilization, transfer, pregnancy, and implantation rates were recorded in each category. Sperm were tested with antibodies for detection of the of the sperm acrosome. MAIN OUTCOME MEASURE(S): Fertilization, transfer, pregnancy and implantation rates, and percentage of acrosome-reacted cells. RESULT(S): The fertilization rate and statistical evaluation showed differences between morphologically normal and pathological sperm and other groups. The freezing-thawing procedure had no influence on the fertilization of testicular sperm, but epididymal frozen-thawed sperm had a higher fertilization rate. Immunofluorescence proved decreasing sperm quality in all groups compared with the control group. This difference is not manifested in other parameters (transfer, pregnancy, implantation rates). CONCLUSION(S): The spermatozoa with varied semen characteristics and good quality, also detected with specific antibodies, gave the best fertilization rates. The paternal effect is not proved in other parameters.
- MeSH
- akrozom imunologie MeSH
- financování organizované MeSH
- hodnocení výsledků zdravotní péče metody MeSH
- imunoanalýza metody MeSH
- imunokomplex analýza MeSH
- intracytoplazmatické injekce spermie statistika a číselné údaje MeSH
- kultivované buňky MeSH
- lidé MeSH
- mužská infertilita epidemiologie imunologie terapie MeSH
- retrospektivní studie MeSH
- sperma cytologie imunologie MeSH
- těhotenství MeSH
- úhrn těhotenství na počet žen v reprodukčním věku MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- MeSH
- dospělí MeSH
- fertilizace in vitro metody MeSH
- kolagenasy MeSH
- kryoprezervace metody MeSH
- kultivované buňky metody MeSH
- lidé MeSH
- odběr biologického vzorku metody MeSH
- spermatidy MeSH
- spermie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
Cíl studie: Transfer kvalitního embrya v programu asistované reprodukce v případě azoospermies dg. Sertolli cells only syndrome (SCO sy) + maturation arrest (MA), kde v testes byl zaznamenánvýskyt převážně Sertoliho buněk s nepatrným množstvím zárodečných buněk, které prokazovalyzástavu zrání (MA) ve stadiu kulatých spermatid. Histologickým hodnocením byla stanovenahypospermatogeneze gr. 3 (minimum jedna spermatida/vzorek).Typ studie: Kazuistika.Název a sídlo pracoviště: Laboratoř IVF, Iscare IVF, a. s., Oddělení biologie a biochemické fertilizace,Ústav molekulární genetiky AV ČR, Praha.Předmět a metodika studie: K úspěšnému řešení uvedeného typu azoospermie je možné použítimunologické, histologické a kultivační metody. Monoklonální protilátky proti akrosomálnímproteinům lidských spermií umožní pomocí immunofl uorescenčního testu zviditelnit prekurzoryspermií v ejakulátu a odhad spermatogeneze je upřesněn histologickou metodou. Po stanovenístupně spermatogeneze je použita kultivace buněčné suspenze in vitro, která umožní získat spermieči jejich prekurzory použitelné v metodě ISCI.Závěr: Využití uvedených metod a jejich vhodná kombinace zvýšila pravděpodobnost zisku kvalitníhoembrya v programu asistované reprodukce.
Objective: The transfer of good quality embryo in the program of assisted reproduction in thecase of azoospermia, dg. Sertolli cells only syndrome (SCO sy) + maturation arrest (MA). Testeswere assessed and found to have a high occurrence of Sertolli cells and very low occurrence ofgerminal cells, which were arrested at the round spermatid level. The histological evaluation washypospermatogenesis gr. 3 (minimum 1 spermatid/sample).Design: Case report.Setting: Laboratory IVF, Iscare, a. s., Department of Biology and Biochemistry of Fertilization,Institute of Molecular Genetics, Czech Academy of Sciences, Prague.Subject and Method: The successful integration of three methods provides a solution for this caseof azoospermia. Immunology and histology can more exactly diagnose the degree of azoospermia.Detection and visualisation of spermatids using monoclonal antibodies against sperm proteinspredicts the eventual occurrence of spermatogenesis, and histological evaluation confi rms theseimmunological fi ndings. Using the information of both methods it is possible to use special invitro cultivation of testicular cells and so obtain injectable spermatozoa, or precursors of sperm,for the ICSI method.Conclusion: The probability of acquisition of goood-quality embryo in the program of assistedreproduction is higher when these three methods are applied in combination.
- MeSH
- asistovaná reprodukce metody MeSH
- finanční podpora výzkumu jako téma MeSH
- lidé MeSH
- mužská infertilita diagnóza etiologie terapie MeSH
- Sertoliho buňky MeSH
- spermatidy MeSH
- spermatogeneze MeSH
- testis anatomie a histologie patologie MeSH
- zrání spermie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- přehledy MeSH
- srovnávací studie MeSH
