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Derivation and characterization of human embryonic stem cell lines from poor quality embryos

Journal of Genetics and Genomics 2009年第4期36卷 229-239页

作者： Weiqiang Liu, Yifei Yin, Xiaolin Long, Yumei Luo, Yonghua Jiang, Wenhong Zhang, Hongzi Du, Shaoying Li, Yuhong Zheng, Qing Li, Xinjie Chen, Baoping Liao, Guohong Xiao, Weihua Wang, Xiaofang Sun Guangzhou Key Laboratory of Reproductive and Genetics Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou 510150, China Institute of Gynecology and Obstetrics The Third Affiliated Hospital of Guangzhou Medical College Guangzhou Key Laboratory of Reproductive and Genetics Guangzhou 510150

Poor quality embryos discarded from in vitro fertilization （IVF） laboratories are good sources for deriving human embryonic stem cell （hESC） lines. In this study, 166 poor quality embryos donated from IVF centers on day 3 were cultured in a blastocyst medium for 2 days, and 32 early blastocysts were further cultured in a blastocyst optimum culture medium for additional 2 days so that the inner cell masses （ICMs） could be identified and isolated easily. The ICMs of 17 blastocysts were isolated by a mechanical method, while those of the other 15 blastocysts were isolated by immunosurgery. All isolated ICMs were inoculated onto a feeder layer for subcultivation. The rates of ICM attachment, primary ICM colony formation and the efficiency of hESC derivation were similar between the ICMs isolated by the two methods （P〉0.05）. As a result, four new hESC lines were established. Three cell lines had normal karyotypes and one had an unbalanced Robertsonian translocation. All cell lines showed normal hESC characteristics and had the differentiation ability. In conclusion, we established a stable and effective method for hESC isolation and culture, and it was confirmed that the mechanical isolation was an effective method to isolate ICMs from poor embryos. These results further indicate that hESC lines can be derived from poor quality embryos discarded by IVF laboratories.

关键词： embryonic stem ceils poor quality embryos inner cell mass mechanical isolation immunosurgery

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