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Methods for derivation of human embryonic stem cells

Cited 59 time in Web of Science Cited 73 time in Scopus
Authors
Kim, Hee Sun; Oh, Sun Kyung; Park, Yong Bin; Ahn, Hee Jin; Sung, Ki Cheong; Kang, Moon Joo; Lee, Lim Andrew; Suh, Chang Suk; Kim, Seok Hyun; Kim, Dong-Wook; Moon, Shin Yong
Issue Date
2005-07-30
Publisher
AlphaMed Press
Citation
Stem Cells. 2005 Oct;23(9):1228-33. Epub 2005 Jul 28.
Keywords
Blastocyst/*cytology/physiologyCell LineCulture MediaEmbryo Culture Techniques/*methodsEmbryo, Mammalian/cytologyHumansPluripotent Stem Cells/*cytology/physiology
Abstract
The expanded blastocysts, developed from 2PN-stage embryos, are generally divided into three categories: a good blastocyst containing a large and distinguishable inner cell mass (ICM), a blastocyst with a small and distinct ICM, and a blastocyst with a poorly defined ICM. In this study, we introduce methods for the derivation of human embryonic stem cells (hESCs) depending on the quality of the blastocysts. An immunosurgical method was used for the good expanded blastocysts. This method, however, raises the probability of ICM loss in cases of hESC derivation from blastocysts with smaller or indistinct ICMs. Furthermore, this method is also associated with a risk of the contamination of the hESCs with animal pathogens. To overcome these shortcomings, the partial- or whole-embryo culture method was used. For blastocysts with no visible ICM, the whole-embryo culture method was used to establish hESCs via the seeding of the entire blastocyst without its zona pellucida directly on a STO feeder layer. However, trophectodermal overgrowth tends to hinder the expansion of the ICM during the initial steps of hESC derivation. Therefore, the partial-embryo culture method was developed to establish hESCs from blastocysts with smaller ICMs. The surgical isolation of the region containing the ICM with an ultra-fine glass pipette alleviates trophectoderm overgrowth. This method is also applicable to blastocysts with large and distinct ICMs, and the efficiency of this method is comparable to that of the immunosurgical method.
ISSN
1066-5099 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16051988

http://hdl.handle.net/10371/39193
DOI
https://doi.org/10.1634/stemcells.2004-0296
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College of Medicine/School of Medicine (의과대학/대학원)Molecular and Genomic Medicine (분자유전체의학전공)Journal Papers (저널논문_분자유전체의학전공)
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