Delimitreva S M
Human IVF Laboratory, Department of Biology, Medical Faculty, Sofia, Bulgaria.
Folia Biol (Praha). 2002;48(4):149-53.
The data about the relation and succession of blastomere fragmentation, cleavage rate and chromatin anomalies in preimplantation mammalian embryos are empirical and controversial at present. In this work we studied the proportion of nuclear fragmentation and condensation in 3-5-cell stage human embryos with no or minimal blastomere fragmentation (morphological class A and B, respectively) and the possibilities to perform FISH chromosomal analyses with them. We observed different stages of chromatin damage in blastomere nuclei corresponding to the steps of nuclear apoptotic changes well known in many cell types. The ploidity analysis of chromosomes 1, 5, 19 and X was determined as successful in embryos which had at least 2 out of 3, 3 out of 4 or 3 out of 5 normal nuclei with an equal number of FISH signals. There was no difference in the percentage of abnormal nuclei among the A- and B-class embryos. Tendencies noted by us suggest that the minimal blastomere fragmentation (up to 20% of perivitelline space) does not preclude the normal nuclear status allowing successful ploidy testing. The presence of condensed chromatin is a critical factor for interphase cytogenetic analysis of single early blastomeres.
目前,关于植入前哺乳动物胚胎中卵裂球碎片化、分裂率和染色质异常之间的关系及连续性的数据是基于经验的,且存在争议。在这项研究中,我们研究了3至5细胞期人类胚胎中核碎片化和凝聚的比例,这些胚胎没有或仅有最小程度的卵裂球碎片化(分别为形态学A类和B类),以及对它们进行荧光原位杂交(FISH)染色体分析的可能性。我们观察到卵裂球细胞核中染色质损伤的不同阶段,这些阶段与许多细胞类型中熟知的核凋亡变化步骤相对应。对于染色体1、5、19和X的倍性分析,在至少3个核中有2个、4个核中有3个或5个核中有3个具有相等数量FISH信号的正常核的胚胎中被确定为成功。A类和B类胚胎中异常核的百分比没有差异。我们注意到的趋势表明,最小程度的卵裂球碎片化(占透明带周隙的20%以下)并不排除正常的核状态,从而允许进行成功的倍性检测。凝聚染色质的存在是对单个早期卵裂球进行间期细胞遗传学分析的关键因素。