Author/Editor		Čižek-Sajko, Mojca
Title		Jedrne nepravilnosti nedozorelih in neuspešno oplojenih človeških jajčnih celic po zorenju v pogojih in vitro
Translated title		Nuclear abnormalities of immature and unsuccessfully fertilized human oocytes after in vitro maturation
Type		monografija
Place		Ljubljana
Publisher		Medicinska fakulteta
Publication year		2007
Volume		str. 95
Language		slo
Abstract		Background: the aim of the study was to determine whether an immature oocyte obtained from a follicle before the dominance phase could be clinically useful in an in vitro fertilization (IVF) procedure in patients with polycystic ovary syndrome (PCOS) stimulated with follitropin (follicle stimulating hormone, FSH) or human chorionic gonadotrophin (hCG). Immature oocytes were cultured in vitro in a complex medium supplemented with gonadotrophins. In immature and unsuccessfully fertilized oocytes, the configuration of chromatin and pertaining microtubules was analyzed. Methods: Sixty IVF procedures with in vitro maturation of oocyes were carried out in 45 infertile couples and analyzed. Only women with POS were included in the study. We compared the success following two different hormone priming protocols: hormone priming with FSH (Group A, n=22) and hormone priming with hCG (Group B, n=38). Immature oocytes were cultured for 24-30 hours in a complex medium supplemented with gonadotrophins. Success was evaluated on the basis of the ability of oocytes to mature, fertilize and develop into embryos, and on the basis of the quality of embryos and their ability to implant in the uterus. Immature and unfertilized oocytes were fixed in paraformaldehyde. After fixation chromatin and microtubules were stained according to Hoechst 33258 and FITC immunolabelling. The configuration of chromatin and pertaining microtubiles was analyzed using immunofluorescence microscopy. Results: The oocyte maturatuon rate, fertilization rate and the embryo cleavage rate were as follows: in Group A 57.6 %, 66.2 % and 90.0 % and in Group B 58.0 %, 66.2 % and 90.3 % (the differences between group were not statistically significant). The quality of embryos evaluated on the basis of multinucleated blastomere presence differed between the groups (P<0.001). (Abs. trunc. at 2000 ch.)
Summary		Izhodišča: Namen raziskave je bil ugotoviti, ali je nezrela jajčna celica iz folikla, ki je razvojno pred izbiro dominantnega folikla, pri preiskovankah s sindromom policističnih ovarijev, spodbujanih s folitropinom ali s humanim horionskim gonadotropinom, lahko klinično uporabna v postopku oploditve z biomedicinsko pomočjo. Nezrele jajčne celice smo zoreli v kompleksnem gojišču z dodarkom gonadotropinov v pogojih in vitro. Pri nedozorelih in neuspešno oplojenih jajčnih celicah smo ugotavljali konfiguracijo kromatina in pripadajočih mikrotubulov. Metode dela: Pri 45 neplodnih parih smo analizirali 60 postopkov zunajtelesne oploditve z zorenjem jajčnih celic v pogojih in vitro. V raziskavo smo vključili le preiskovanke s sindromom policističnih jajčnikov (PCOS). Na punkcijo antralnih foliklov smo jih pripravili s hormonskim spodbujanjem po dveh različnih protokolih: hormonsko spodbujanje rasti foliklov s folitropinom (folikle spodbujajočim hormonom, FSH) (skupina A, n=22) in hormonsko spodbujanje ovulacije s humanim horionskim gonadotropinom (hCG) (skupina B, n=38). Nezrele jajčne celice smo kultivirali 24-30 uur v kmpleksnem gojišču z dodatkom gonadotropinov. Primerjali smo uspešnost obeh protokolov, ki smo jih ovrednotili s sposobnostjo jajčnih celic za zorenje, za oploditev in za razvoj v zarodke, ter s kakovostjo zarodkov in njihovo sposobnostjo za ugnezditev v maternico. Nedozorele in neuspešno oplojene jajčne celice smo fiksirali v paraformaldehidu ter jim z imunofluorescentnimi barvili Hoechst 33258 in FITC obarvali kromatin in označili tubulin. Z imunofluorescentno mikroskopijo smo nato ocenjevali konfiguracijo kromatina in pripadajočih mikrotubulov. Rezultati: Stopnja dozorevanja jajčnih celic, stopnja oploditve in stopnja delitve zigot v zarodke se med skupinama A in B ni razlikovala: pri preiskovankah skupine A je bila 57.6 %, 66.2 % in 90.0 %, pri preiskovankah skupine B pa 58.0 %, 66.2 % in 90.3 %. (Izvl. skrajšan na 2000 zn.)
Descriptors		POLYCYSTIC OVARY SYNDROME GONADOTROPINS OOCYTES CHROMATIN MICROTUBULES FERTILIZATION IN VITRO EMBRYO PREGNANCY RATE COHORT STUDIES