【摘要】 在体外受精过程中,通过胚胎植入前遗传性诊断(PGD)对有遗传风险患者的胚胎进行植入前活检和遗传学分析,选择无遗传性疾病的胚胎植入子宫,而PGD诊断异常的胚胎则会被丢弃。本研究尝试将PGD异常胚胎用于分离人胚胎干细胞,以获得携带遗传缺陷的人胚胎干细胞系。利用荧光原位杂交技术对第3-5天胚胎进行PGD检测,结果异常的胚胎进一步用于分离获取胚胎干细胞系,然后对h ES细胞系进行核型及干细胞表面标记、多能性基因表达、端粒酶活性以及分化能力等特征性鉴定。总共从13个PGD异常胚胎中分离获得8个人胚胎干细胞系,建系效率为61.5%,其中1个核型正常,5个核型异常。说明利用PGD异常胚胎可以获得携带遗传缺陷的人胚胎干细胞系,不仅为评估PGD技术临床结论的准确性提供了一种新方法,更重要的是为研究各种遗传性疾病的发病机理提供了有效的细胞模型。更多还原

【Abstract】 In the process of in vitro fertilization,the embryos from patients with genetic risk will take biopsy and the pre-implantation genetic diagnosis(PGD). The embryologist will select normal embryos for the reproductive purpose,while abnormal embryos with genetic mutation will be discarded. This study attempted to isolate human embryonic stem cell(h ESC) lines with genetic defects from PGD-analyzed abnormally embryos. The day 3-5 embryos were biopsied for PGD analysis with fluorescence in situ hybridization(FISH),and abnormal PGD-analyzed embryos were further used for derivation of h ES cell lines. The h ESC lines were further characterized with the expression of markers and pluripotencyrelated genes,telomerase activity,karytping and differentiation activity in vitro. Totally 8 h ESC lines were isolated from13 abnormal PGD-analyzed embryos,and the isolation rate was 61. 5%. Among them,one was normal and the other 7ones with abnormal karyotype. It suggested that h ESC lines with genetic defects can be derived from abnormal PGD-analyzed embryos. Such h ESC lines will provide a new approach to assess the accuracy of PGD results and a powerful tool for the study of human genetic disorders.更多还原