Shkumatov Artem, Kuznyetsov Valeriy, Cieslak Jeanine, Ilkevitch Yuri, Verlinsky Yury
Reproductive Genetics Institute, 2825 North Halsted Street, Chicago, IL 60657, USA.
Reprod Biomed Online. 2007 Apr;14(4):498-503. doi: 10.1016/s1472-6483(10)60899-1.
It has previously been shown that it is possible to obtain metaphase chromosomes from single blastomeres converted into metaphase in the cytoplasm of a mouse zygote. This method is highly labour intensive and cannot be performed outside the preimplantation genetic diagnosis (PGD) laboratory, so to overcome these limitations, a method was developed for obtaining metaphase spreads from single biopsied blastomeres using different chemicals. The substances tested were calyculin A, caffeine, paclitaxel and colcemid in a total of 496 disaggregated and 234 biopsied blastomeres from day 3 embryos. It was demonstrated that the optimal method involved a combined use of 'selective biopsy' (selection of the biopsied blastomere according to morphological criteria) and exposure to caffeine. This resulted in shortening the mean incubation time of biopsied blastomeres, with a metaphase formation rate of 80%. The method is simple for obtaining metaphases from single blastomeres, and may be implemented in clinical practice of PGD for structural rearrangements.
先前已经表明，从在小鼠受精卵细胞质中转化为中期的单个卵裂球获得中期染色体是可能的。这种方法劳动强度极大，且不能在植入前基因诊断（PGD）实验室之外进行，因此为克服这些限制，开发了一种使用不同化学物质从单个活检卵裂球获得中期染色体铺展的方法。所测试的物质为calyculin A、咖啡因、紫杉醇和秋水仙酰胺，共对496个解离的和234个来自第3天胚胎的活检卵裂球进行了测试。结果表明，最佳方法是“选择性活检”（根据形态学标准选择活检卵裂球）和暴露于咖啡因的联合使用。这使得活检卵裂球的平均孵育时间缩短，中期形成率为80%。该方法从单个卵裂球获得中期染色体很简单，可用于PGD临床实践中对结构重排的检测。
