Vajta G, Lewis I M, Hyttel P, Thouas G A, Trounson A O
Centre for Early Human Development, Monash Institute of Reproduction and Development, Monash University, Clayton, Victoria, Australia.
Cloning. 2001;3(2):89-95. doi: 10.1089/15204550152475590.
Until now, micromanipulators have been regarded as indispensable for somatic cell nuclear transfer. This paper describes an improved zona-free nuclear transfer procedure with manual bisection of oocytes, selection of cytoplasts by Hoechst staining, and two-step fusion of somatic cells from primary granulosa cell cultures with two cytoplasts. Blastocyst rates in the three systems tested for zona-free embryo culture were 0%, 18%, and 36% for microdrops, well of the wells (WOW system), and microcapillaries (GO system), respectively. This simple, rapid, and inexpensive procedure may become a useful alternative to the existing techniques for somatic cell nuclear transfer for large-scale application of the technology.
到目前为止，显微操作仪一直被视为体细胞细胞核移植不可或缺的工具。本文描述了一种改进的无透明带核移植程序，包括手动对卵母细胞进行二分、通过Hoechst染色选择细胞质体，以及将来自原代颗粒细胞培养物的体细胞与两个细胞质体进行两步融合。在用于无透明带胚胎培养的三个系统中，微滴、孔中孔（WOW系统）和微毛细管（GO系统）的囊胚率分别为0%、18%和36%。这种简单、快速且廉价的程序可能成为现有体细胞细胞核移植技术的一种有用替代方法，以便该技术能够大规模应用。
