27 KB209 KB26 KB192920262002Kovačič, BorutVlaisavljević, Veljkoletnik:71str. I-13-I-17številka:supl. 1ISSN:1318-0347COBISSID:857151URN:URN:NBN:SI:doc-GWAW3DSVslSlovensko zdravniško društvoZdravniški vestnikfertilizationFertilization in vitrohumana reprodukcijain vitro fertilizationJajčna celicaMeiosismejozaOocyteoploditevOploditev in vitroRedukcijska delitevRezultati in vitro zorenja mejotsko nezrelih človeških jajčnih celic v enostavnem gojišču| Results of in vitro maturation of meiotically immature human oocytes in a simple medium|Background. Among oocytes obtained during aspiration of preovulatory ovarian follicles in hormonally stimulated cycles, we ascertained the percentage of immature oocytes with the nucleus in the metaphase (M I oocytes) or even in the prophase (GV oocytes) of the first meiotic division and their capacity to mature in vitro in a simple medium without hormonal supplements. Methods. In 818 women, stimulated gonadotropin releasing hormone agonist (GnRHa) and gonadotropins, aspiration of preovulatory size follicles yielded 4972 oocytes. From these we denuded cells of cumulus oophorus and corona, meiotic maturity was evaluated under a microscope. Cells in the metaphase of the second meiotic division ( M II oocytes) and those maturing after 5 hours were used clinically in the intracytoplasmic sperm injection (ICSI) procedure. Immature cells were left in the simple medium. The degree of their nuclear maturity was evaluated after one and after two days of culture. In vitro maturation was clinically used also in 14 cycles with no mature oocytes. Results. Among 4731 oocytes with denuded corona and culumus, 4199 (88.8%) were mature M II oocytes, 295 (6.2%) immature M I oocytes and 237 (5%) immature GV oocytes. Under in vitro conditions, 68.7% (90/131) GV oocytes attained maturity. Among M I oocytes, 63.6% (136/214) cells matured already after 5 hours and 26.6% (57/214) until the next day. In all 14 women with only immature oocytes, the embryos for embryotransfer were obtained after in vitro maturation and ICSI procedure. Theresults was four pregnancies and two deliveries. Conclusions. Immature oocytes, obtained in hormonally stimulated cycles, may become clinically applicable if left to mature in vitro in a simple medium without supplementation of growth factors and hormonesIzhodišča. V raziskavi ugotavljamo, kolikšen je med jajčnimi celicami, ki jih dobimo pri aspiraciji preovulatornih jajčnih foliklov v hormonsko spodbujanih ciklih, delež nezrelih jajčnih celic z jedrom v metafazi (M I oociti) ali celo profazi (GV oociti) prve redukcijske delitve in kakšna je sposobnost zorenja jajčnih celic in vitro v preprostem gojišču brez dodatnih hormonov. Metode. Pri 818 ženskah, spodbujanih z agonisti sproščevalcev gonadotropinov, smo po aspiraciji preovulacijskih jajčnih foliklov dobili 4972 jajčnih celic. Le-tem smo odstranili celice kumulusa ooforusa in korone radiate in pod mikroskopom ocenili mejotsko zrelost. Celice v metafazi druge redukcijske delitve (M II oociti) in tiste, ki so dozorele po 5 urah, smo uporabili klinično v postopku intracitoplazmatskega injiciranja semenčice v jajčno celico (ICSI). Preostale nezrele celice smo pustili v enostavnem gojišču. Stanje njihove jedrne zrelosti smo ocenjevali po enem oziroma dveh dnevih zorenja. In vitro zorenje jajčnih celic smo klinično uporabili tudi v 14 ciklih z vsemi nezrelimi celicami. Rezultati. Med 4731 jajčnimi celicami z odstranjeno korono in kumulusom je bilo 4199 (88,8%) zrelih M II oocitov, 295 (6,2%) nezrelih M I oocitov in 237 (5%) nezrelih GV oocitov. V pogojih in vitro je dozorelo 68,7% (90/131) GV oocitov. Med M I oociti je 63,6% (136/214) celic dozorelo že po 5 urah in 26,6% (57/214) do naslednjega dne. Pri vseh 14 ženskah s samo nezrelimi jajčnimi celicami na dan aspiracije smo po zorenju in vitro in ICSI postopku pridobili zarodke za prenos. Dosegli smo 4 nosečnosti in dve rojstvi. Zaključki. Nezrele jajčne celice, ki jih dobimo v hormonsko spodbujanih ciklih, lahko postanejo klinično uporabne, če jih pustimo zoreti in vitro v enostavnem gojišču brez dodanih rastnih faktorjev in hormonovTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaSlovensko zdravniško društvo