Serial Pronuclear Transfer Increases the Developmental Potential of In Vitro-Matured Oocytes in Mouse Cloning1
Serial Pronuclear Transfer Increases the Developmental Potential of In Vitro-Matured Oocytes in Mouse Cloning1
复制标题
连续原核移植提高小鼠克隆体外成熟卵母细胞的发育潜力1
作者：
B. Heindryckx;A. Rybouchkin;J. Van der Elst;M. Dhont
摘要在动物和人的核移植实验中，体外成熟的生殖囊泡卵母细胞是一种有趣的胞质受体来源。我们研究了两个技术方面，可能会提高体外成熟的萌发囊泡卵母细胞构建的核移植小鼠胚胎的发育潜力。在第一步中，比较了两种成熟培养基对源自体外成熟卵母细胞的NT胚胎的胚胎发育的影响。补充血清和促性腺激素的卵母细胞成熟培养液提高了NT胚胎的发育率构建从体外成熟的卵母细胞，但它仍然不如在体内成熟的中期II（MII）的卵母细胞。第二，我们研究了从NT合子来源于体外和体内成熟的卵母细胞在体内受精的合子胞质的连续原核转移的效果。通过使用差异染色计数滋养外胚层和内细胞团细胞的细胞核来评价囊胚质量。序贯原核移植显着提高胚泡形成率NT胚胎起源于在体外成熟的卵母细胞获得的速度与在体内成熟的MII卵母细胞。我们的结论是，NT胚胎的发育潜力构建从体外成熟的卵母细胞可以优化系列原核转移到体内产生的合子胞质。
Abstract In vitro-matured germinal vesicle oocytes are an interesting source of cytoplast recipients in both animal and human nuclear transfer (NT) experiments. We investigated two technical aspects that might improve the developmental potential of nuclear transfer mouse embryos constructed from in vitro-matured germinal vesicle oocytes. In a first step, the effect of two maturation media on the embryonic development of NT embryos originating from in vitro-matured oocytes was compared. Supplementation of the oocyte maturation medium with serum and gonadotrophins improved the developmental rate of NT embryos constructed from in vitro-matured oocytes, but it was still inferior to that obtained with in vivo-matured metaphase II (MII) oocytes. Second, we investigated the effect of serial pronuclear transfer from NT zygotes originating from both in vitro- and in vivo-matured oocytes to in vivo-fertilized zygotic cytoplasts. Blastocyst quality was evaluated by counting nuclei from trophectoderm and inner cell mass cells using a differential staining. Sequential pronuclear transfer significantly improved the blastocyst formation rate of NT embryos originating from in vitro-matured oocytes up to the rate obtained with in vivo-matured MII oocytes. We conclude that the developmental potential of NT embryos constructed from in vitro-matured oocytes can be optimized by serial pronuclear transfer to in vivo-produced zygotic cytoplasts.
影响因子：
3.6
作者：
ERBACH, GT;LAWITTS, JA;BIGGERS, JD
通讯作者：
BIGGERS, JD