Биотехнологические основы получения клонированных эмбрионов свиней

Термин «клон» в биотехнологии животных обозначает организм, полученный в результате неполового размножения, который одновременно является прямым потомком и генетической копией родительского организма. На сегодняшний день домашняя свинья (Sus scrofa domestica) представляется наиболее интересным объектом в исследованиях по клонированию. Клонирование свиней имеет широкий спектр потенциальных возможностей использования в различных областях научной и хозяйственной деятельности человека. Тем не менее эффективность получения клонированных эмбрионов свиней все еще остается ниже, чем других видов сельскохозяйственных животных, в частности лошадей и крупного рогатого скота. Соматическое клонирование – сложная многоступенчатая технология, на каждом этапе которой более восприимчивые к изменениям окружающих условий ооциты свиней испытывают неблагоприятные воздействия различных по своей природе факторов (механические, физические, химические). На этапе созревания ооцитов происходят изменения клеточных ультраструктур ооплазмы, которые играют важную роль в последующем репрограммировании ядра пересаженной донорской клетки. Донорские соматические клетки перед переносом в ооцит синхронизируют в стадии G0/G1 клеточного цикла с целью обеспечения нормальной плоидности клонированного эмбриона. При удалении ядра у созревших in vitro ооцитов свиней следует обращать внимание на проблему сохранения жизнеспособности клеток после извлечения собственного ядерного материала. В ходе реконструирования соматическую клетку с помощью микроинструментов помещают в перивителлиновое пространство, где ранее находилось первое полярное тельце, или в цитоплазму энуклеированного ооцита. Метод ручного клонирования (handmade cloning) предполагает удаление ядра ооцита с последующим слиянием с донорской клеткой без помощи микроманипуляционной техники. Повышенная чувствительность ооцитов к факторам окружающей среды обусловливает особые требования к выбору системы in vitro культивирования клонированных эмбрионов свиней. В рамках настоящего обзора проведен мониторинг современных методов, используемых при получении клонированных эмбрионов, выявлены технологические особенности, препятствующие повышению эффективности метода соматического клонирования свиней.

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