Eglitis M A
J Exp Zool. 1980 Aug;213(2):309-13. doi: 10.1002/jez.1402130222.
A rapid method to produce tetraploid embryos by blastomere fusion using polyethylene glycol (PEG) has been developed. Individual four-cell stage blastomeres were aggregated into pairs with phytohemagglutinin (PHA) and then treated with 45% (w/v) PEG (MW 1,000). 56.8% (804/1,416) of treated blastomere pairs fused. Tetraploid blastomeres were aggregated into quartets or doublets to restore cell number equivalent to that of whole or half diploid embryos, respectively. Best development was obtained with quartets, 72.9% (78/107) of which cavitated. 46.4% (13/28) of doublets and 31.7% (13/41) of singly cultured tetraploid blastomeres formed cavitated structures. Uniform tetraploidy was confirmed by fixing cavitated embryos and analyzing their metaphase plates. The fusion method described here replaces Sendai virus with a chemically defined fusogen and confirms earlier observations (Snow, '73; Graham, '71) that tetraploidy does not prohibit preimplantation development. This fusion method obviates some of the drawbacks of cytochalasin B-induced tetraploidy, namely the blocking of cleavage and the potential of diploid/tetraploid mosaicism.
一种利用聚乙二醇（PEG）通过卵裂球融合产生四倍体胚胎的快速方法已被开发出来。将单个四细胞期的卵裂球与植物血凝素（PHA）聚集成对，然后用45%（w/v）的PEG（分子量1000）处理。56.8%（804/1416）的处理过的卵裂球对发生融合。四倍体卵裂球被聚集成四重体或双联体，以分别恢复相当于完整或半个二倍体胚胎的细胞数量。四重体的发育效果最佳，其中72.9%（78/107）形成了囊胚腔。双联体的46.4%（13/28）和单独培养的四倍体卵裂球的31.7%（13/41）形成了有囊胚腔结构。通过固定有囊胚腔的胚胎并分析其中期板，证实了均匀的四倍体状态。这里描述的融合方法用化学定义的融合剂取代了仙台病毒，并证实了早期的观察结果（斯诺，1973年；格雷厄姆，1971年），即四倍体并不妨碍植入前的发育。这种融合方法消除了细胞松弛素B诱导四倍体的一些缺点，即卵裂的阻断和二倍体/四倍体嵌合体的可能性。
