Xinjie Chen1, Yumei Luo, Yong Fan, Lei Yue, Xueshi Wu, Yaoyong Chen, Xiaofang Sun. 1. Key Laboratory for Major Obstetric Diseases of Guangdong Province, Guangzhou Key Laboratory of Reproduction and Genetics, Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Duobao Road 63#, Guangzhou, Guangdong, 510150, People's Republic of China. lucychen23@yahoo.cn
Abstract
PURPOSE: Human embryonic stem cells (hESCs) are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. The need for new hESC lines motivated our attempts to find a new resource for the derivation of hESC lines. The aim of this work was to establish more hESC lines from abnormal fertilized zygotes and to meet the emerging requirements for their use in cell replacement therapies, disease modeling, and basic research. METHODS: A total of 130 tripronuclear human zygotes was collected 18-20 h post-insemination and cultured in a modified culture medium. The inner cell mass of 12 blastocysts were isolated by a mechanical method in order to establish embryonic stem cell lines. RESULTS: We established four hESC lines derived from 130 trinuclear zygotes, one of which was triploid and the others were diploid. The efficiency of deriving hESC lines is 3.08 %. The ratio of deriving triploid and diploid hESC lines is 1:3. All of these hESC lines exhibited similar markers of undifferentiated hESCs and had the typical morphology of hESCs, a capacity for long-term proliferation, and pluripotent differentiation potential both in vivo and in vitro. CONCLUSIONS: These abnormal zygotes, which otherwise would have been discarded, can serve as an alternative source for normal euploid hESC lines.
PURPOSE:Human embryonic stem cells (hESCs) are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. The need for new hESC lines motivated our attempts to find a new resource for the derivation of hESC lines. The aim of this work was to establish more hESC lines from abnormal fertilized zygotes and to meet the emerging requirements for their use in cell replacement therapies, disease modeling, and basic research. METHODS: A total of 130 tripronuclear human zygotes was collected 18-20 h post-insemination and cultured in a modified culture medium. The inner cell mass of 12 blastocysts were isolated by a mechanical method in order to establish embryonic stem cell lines. RESULTS: We established four hESC lines derived from 130 trinuclear zygotes, one of which was triploid and the others were diploid. The efficiency of deriving hESC lines is 3.08 %. The ratio of deriving triploid and diploid hESC lines is 1:3. All of these hESC lines exhibited similar markers of undifferentiated hESCs and had the typical morphology of hESCs, a capacity for long-term proliferation, and pluripotent differentiation potential both in vivo and in vitro. CONCLUSIONS: These abnormal zygotes, which otherwise would have been discarded, can serve as an alternative source for normal euploid hESC lines.
Authors: A H Sathananthan; J J Tarin; L Gianaroli; S C Ng; V Dharmawardena; M C Magli; R Fernando; A O Trounson Journal: Hum Reprod Update Date: 1999 Sep-Oct Impact factor: 15.610
Authors: Chad A Cowan; Irina Klimanskaya; Jill McMahon; Jocelyn Atienza; Jeannine Witmyer; Jacob P Zucker; Shunping Wang; Cynthia C Morton; Andrew P McMahon; Doug Powers; Douglas A Melton Journal: N Engl J Med Date: 2004-03-03 Impact factor: 91.245