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与体外成熟的牛卵母细胞融合后,在单个人类卵裂球中快速观察中期染色体。

Rapid visualization of metaphase chromosomes in single human blastomeres after fusion with in-vitro matured bovine eggs.

作者信息

Willadsen S, Levron J, Munné S, Schimmel T, Márquez C, Scott R, Cohen J

机构信息

Institute for Reproductive Medicine and Science of Saint Barnabas, Livingston, New Jersey 07052, USA.

出版信息

Hum Reprod. 1999 Feb;14(2):470-5. doi: 10.1093/humrep/14.2.470.

Abstract

The present study was aimed to facilitate karyotyping of human blastomeres using the metaphase-inducing factors present in unfertilized eggs. A rapid technique for karyotyping would have wide application in the field of preimplantation genetic diagnosis. When cryopreserved in-vitro matured bovine oocytes were fused with human blastomeres, the transferred human nuclei were forced into metaphase within a few hours. Eighty-seven human blastomeres from abnormal or arrested embryos were fused with bovine oocytes in a preclinical study. Fusion efficiency was 100%. In 21 of the hybrid cells, no trace of human chromatin was found. Of the remaining 66, 64 (97%) yielded chromosomes suitable for analysis. The method was used to karyotype embryos from two patients with maternal translocations. One embryo which was judged to be karyotypically normal was replaced in the first patient, resulting in one pregnancy with a normal fetus. None of the second patient's embryos was diagnosed as normal, and hence none was transferred. The results of the present study demonstrated that the ooplasmic factors which induce and maintain metaphase in bovine oocytes can force transferred human blastomere nuclei into premature metaphase, providing the basis for a rapid method of karyotyping blastomeres from preimplantation embryos and, by implication, cells from other sources.

摘要

本研究旨在利用未受精卵中存在的中期诱导因子促进人类卵裂球的核型分析。一种快速的核型分析技术将在植入前基因诊断领域有广泛应用。当将冷冻保存的体外成熟牛卵母细胞与人类卵裂球融合时,转移的人类细胞核在数小时内被强制进入中期。在一项临床前研究中,将来自异常或发育停滞胚胎的87个人类卵裂球与牛卵母细胞融合。融合效率为100%。在21个杂交细胞中,未发现人类染色质的踪迹。在其余66个细胞中,64个(97%)产生了适合分析的染色体。该方法被用于对两名患有母源易位的患者的胚胎进行核型分析。第一个患者中一个被判定核型正常的胚胎被移植,结果一次妊娠并产下一个正常胎儿。第二个患者的胚胎均未被诊断为正常,因此均未进行移植。本研究结果表明,牛卵母细胞中诱导和维持中期的卵质因子可迫使转移的人类卵裂球核进入早熟中期,为一种快速分析植入前胚胎卵裂球核型的方法提供了基础,并且由此也为分析其他来源的细胞提供了基础。

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