Obtaining metaphase spreads from single blastomeres for PGD of chromosomal rearrangements

REPRODUCTIVE BIOMEDICINE ONLINE

卷:14

Article

Shkumatov, Artem ; Kuznyetsov, Valeriy ; Cieslak, Jeanine ; Ilkevitch, Yuri ; Verlinsky, Yury

关键词: blastomeres;    caffeine;    chromosomal analysis;    chromosomal rearrangements;    metaphase spreads;    preimplantation genetic diagnosis;   

DOI  :  10.1016/S1472-6483(10)60899-1

来源: Elsevier

【 摘 要 】

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.

【 授权许可】

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