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2015 | 11 | 3 |
Tytuł artykułu

Iniekcja plemnika do cytoplazmy (ICSI) jako alternatywa dla standardowego zapłodnienia in vitro u świń

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Treść / Zawartość
Warianty tytułu
EN
Intra-cytoplasmic sperm injection (ICSI) as an alternative to standard in vitro fertilization in pigs
Języki publikacji
PL
Abstrakty
PL
Iniekcja plemnika do cytoplazmy oocytu (ICSI) jest jedną z metod wspomaganego rozrodu, która znalazła zastosowanie w pozaustrojowej produkcji zarodków u zwierząt oraz leczeniu niepłodności u ludzi. Metoda ICSI umożliwia eliminację zjawiska polispermii, wykorzystanie plemników jako wektorów egzogennego DNA w celu uzyskania zwierząt transgenicznych, wspomaga tworzenie programów zachowania bioróżnorodności oraz poznanie mechanizmów zachodzących w trakcie zapłodnienia i wczesnego rozwoju zarodkowego. Zapłodnieniu metodą ICSI poddaje się oocyty dojrzałe in vitro, in vivo i kriokonserwowane oraz nasienie ejakulowane bądź najądrzowe, świeże lub konserwowane. Iniekcję plemnika przeprowadza się w mikroskopie odwróconym z dołączonym mikromanipulatorem wyposażonym w pipetę podtrzymującą oraz iniekcyjną. Po wykonaniu iniekcji oocyty poddaje się sztucznej aktywacji i przeznacza do hodowli in vitro. Uzyskane zarodki poddaje się ocenie jakości. Pomimo potencjalnych możliwości związanych z ICSI wciąż istnieją problemy, które ograniczają wdrożenie tej metody zapłodnienia do pozaustrojowej produkcji zarodków na skalę komercyjną. Czynnikami mogącymi wpływać na efektywność ICSI są: jakość oocytów i plemników oraz ich przygotowanie do zapłodnienia, uszkodzenia oocytu powstałe w trakcie iniekcji, toksyczność związków wykorzystywanych do spowolnienia ruchu plemników i aktywacja oocytów po iniekcji. W wyniku zapłodnienia metodą ICSI liczba uzyskanych zarodków jest niska, a ich jakość często jest obniżona. Niezbędne są zatem dalsze badania optymalizujące nie tylko samą technikę zapłodnienia, ale również poszczególne jej etapy, tj. dojrzewanie in vitro oocytów oraz przygotowanie plemników.
EN
Injection of sperm into the oocyte cytoplasm (ICSI) is a method of assisted reproduction that has been used for in vitro production of embryos in animals and treatment of infertility in humans. ICSI makes it possible to eliminate polyspermy and to use sperm as vectors of exogenous DNA to obtain transgenic animals. It aids in the creation of biodiversity conservation programmes and in understanding mechanisms taking place during fertilization and early embryonic development. Fertilization by ICSI is performed using IVM, in vivo and cryopreserved oocytes and ejaculated or epididymal semen, fresh or preserved. Sperm injection is carried out using an inverted microscope equipped with a micromanipulator attached to the holding and injection pipettes. After injection, the oocytes are artificially activated and cultured in vitro. The embryos obtained are evaluated. Despite the potential of ICSI, there are still problems limiting its implementation for in vitro production of embryos on a commercial scale. Factors that may influence the efficiency of ICSI are oocyte and sperm quality and their preparation for fertilization, damage to the oocyte during injection, toxicity of compounds used to slow the sperm, and oocyte activation after injection. The number of embryos obtained as a result of ICSI fertilization is low and their quality is often reduced. Further studies are needed to optimize not only the fertilization technique, but also its individual stages, i.e. in vitro maturation of oocytes and sperm preparation.
Wydawca
-
Rocznik
Tom
11
Numer
3
Opis fizyczny
s.55-66,bibliogr.
Twórcy
autor
  • Dział Biotechnologii Rozrodu Zwierząt, Instytut Zootechniki - Państwowy Instytut Badawczy, ul.Krakowska 1, 32-083 Balice k.Krakowa
Bibliografia
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