Document Detail

Human embryos derived by somatic cell nuclear transfer using an alternative enucleation approach.
MedLine Citation:
PMID:  19196043     Owner:  NLM     Status:  MEDLINE    
Somatic cell nuclear transfer (SCNT) was used to generate patient-specific embryonic stem cells (ESCs) from blastocysts cloned by nuclear transfer (ntESCs). In this study, a total of 135 oocytes were obtained from 12 healthy donors (30-35 years). Human oocytes, obtained within 2 h following transvaginal aspiration, were enucleated using a Spindle Imaging System to position the spindle and chromosomes that lay on the metaphase plate, and a Zona Infrared Laser Optical System was used to open a single hole in the zona pellucida at the ~ 2 o'clock position. Human fibroblasts and lymphocytes were used to construct SCNT embryos. Nearly half (26 of 58) of the oocytes were fused after electrofusion and embryo development rates were 96.2% (two-cell, 25 of 26), 92.3% (four-cell, 24 of 26), 61.5% (eight-cell, 16 of 26), 34.6% (16-cell, 9 of 26), 26.9% (morula, 7 of 26), and 19.2% (blastocyst, 5 of 26), respectively, following incubation in improved G-series sequential medium. One cloned blastocyst was used for STR-DNA identification and genetic polymorphism analysis of mtDNA, and STR-DNA analysis of all cloned blastocysts indicated they were derived from SCNT. Quantitative analysis showed that mtDNA of cloned embryos reflected the change tendency of those observed in human IVF embryos. Our research provides an alternative enucleation approach for producing human SCNT-derived blastocysts, and may aid in providing a new method for human therapeutic cloning.
Jianyuan Li; Xuexia Liu; Haiyan Wang; Shouxin Zhang; Fujun Liu; Xuebo Wang; Yanwei Wang
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cloning and stem cells     Volume:  11     ISSN:  1557-7457     ISO Abbreviation:  Cloning Stem Cells     Publication Date:  2009 Mar 
Date Detail:
Created Date:  2009-03-16     Completed Date:  2009-05-26     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101125444     Medline TA:  Cloning Stem Cells     Country:  United States    
Other Details:
Languages:  eng     Pagination:  39-50     Citation Subset:  IM    
Shandong Research Center of Stem Cell Engineering, China.
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MeSH Terms
Blastocyst / cytology,  physiology*
Cloning, Organism / methods*
Fertilization in Vitro
Fibroblasts / cytology,  physiology
Lymphocytes / cytology,  physiology
Nuclear Transfer Techniques*
Oocytes / cytology,  growth & development
Ovulation Induction

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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