Heindryckx B, De Sutter P, Gerris J, Dhont M, Van der Elst J
Infertility Centre, Ghent University Hospital, De Pintelaan 185, 9000 Ghent, Belgium.
Hum Reprod. 2007 Jul;22(7):1982-90. doi: 10.1093/humrep/dem106. Epub 2007 May 18.
Somatic cell nuclear transfer (SCNT) involves the transfer of somatic cell nuclei into enucleated oocytes. Because human in vivo matured oocytes are scarcely available, we investigated whether in vitro matured (IVM) germinal vesicle (GV) oocytes could also support preimplantation development of human cloned embryos.
Three groups were used for SCNT: in vitro matured GV oocytes (IVM oocytes), 'in vivo' matured oocytes (in vivo oocytes) and 'failed fertilized' oocytes after routine-ICSI (FF oocytes). After removal of the chromosome-spindle complex, cumulus cell nuclei were injected, and oocytes were artificially activated and cultured.
In total 61, 54 and 45 metaphase II oocytes were used for SCNT in the three groups, respectively. Survival and pronuclear rates were 59, 78 and 58% and 61, 64 and 50%, respectively. Of the 22 activated IVM oocytes, 13 cleaved to the 2-cell stage, whereby 2 morulae were formed. For the in vivo oocytes, 17 of 27 activated oocytes cleaved to the 2-cell stage and 1 morula was observed. Cleavage to the 2-cell stage in the group of FF oocytes was compromised.
To our knowledge, this is the first report describing development of cloned human embryos using IVM oocytes and non-autologous transfer using a conventional method of SCNT.
体细胞核移植（SCNT）涉及将体细胞核转移到去核卵母细胞中。由于人类体内成熟的卵母细胞很难获得，我们研究了体外成熟（IVM）的生发泡（GV）卵母细胞是否也能支持人类克隆胚胎的植入前发育。
三组用于SCNT：体外成熟的GV卵母细胞（IVM卵母细胞）、“体内”成熟的卵母细胞（体内卵母细胞）和常规卵胞浆内单精子注射（ICSI）后“受精失败”的卵母细胞（FF卵母细胞）。去除染色体-纺锤体复合物后，注射卵丘细胞核，并对卵母细胞进行人工激活和培养。
三组分别有61、54和45个中期II卵母细胞用于SCNT。存活率和原核形成率分别为59%、78%和58%以及61%、64%和50%。在22个激活的IVM卵母细胞中，13个分裂至2细胞期，形成2个桑葚胚。对于体内卵母细胞，27个激活的卵母细胞中有17个分裂至2细胞期，观察到1个桑葚胚。FF卵母细胞组分裂至2细胞期的情况较差。
据我们所知，这是第一份描述使用IVM卵母细胞克隆人类胚胎发育以及使用传统SCNT方法进行非自体移植的报告。
