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Evaluating the genomic and sequence integrity of human ES cell lines; comparison to normal genomes

Title
Evaluating the genomic and sequence integrity of human ES cell lines; comparison to normal genomes
Authors
Walter D. Funk;Ivan Labat;Michael D. West;Rade Drmanac;Kyoung-Jin Shin;Myung Jin Park;Karen B. Chapman;Markus D. Lacher;Lorraine Meisner;Julie A. Johnson;Birgit Stache-Crain;Stacy Caillier;Nadia Sheibani;Daniel Steiger;Elen Rosler;Jorge R. Oksenberg;Pierre-Antoine Gourraud;Janani Sampathkumar
Issue Date
2012
Journal Title
Stem Cell Research
ISSN
1873-5061
Citation
Stem Cell Research, Vol.8(2) : 154~164, 2012
Abstract
Copy number variation (CNV) is a common chromosomal alteration that can occur during in vitro cultivation of human cells and can be accompanied by the accumulation of mutations in coding region sequences. We describe here a systematic application of current molecular technologies to provide a detailed understanding of genomic and sequence profiles of human embryonic stem cell (hESC) lines that were derived under GMP-compliant conditions. We first examined the overall chromosomal integrity using cytogenetic techniques to determine chromosome count, and to detect the presence of cytogenetically aberrant cells in the culture (mosaicism). Assays of copy number variation, using both microarray and sequence-based analyses, provide a detailed view genomic variation in these lines and shows that in early passage cultures of these lines, the size range and distribution of CNVs are entirely consistent with those seen in the genomes of normal individuals. Similarly, genome sequencing shows variation within these lines that is completely within the range seen in normal genomes. Important gene classes, such as tumor suppressors and genetic disease genes, do not display overtly disruptive mutations that could affect the overall safety of cell-based therapeutics. Complete sequence also allows the analysis of important transplantation antigens, such as ABO and HLA types. The combined application of cytogenetic and molecular technologies provides a detailed understanding of genomic and sequence profiles of GMP produced ES lines for potential use as therapeutic agents.
URI
http://www.sciencedirect.com/science/article/pii/S1873506111001231

http://ir.ymlib.yonsei.ac.kr/handle/22282913/91866
DOI
10.1016/j.scr.2011.10.001
Appears in Collections:
1. 연구논문 > 1. College of Medicine > Dept. of Forensic Medicine
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