Trace levels of mitomycin C disrupt genomic integrity and lead to DNA damage response defect in long-term-cultured human embryonic stem ... View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2015-01

AUTHORS

Di Zhou, Ge Lin, Si-Cong Zeng, Bo Xiong, Ping-Yuan Xie, De-Hua Cheng, Qing Zheng, Qi Ouyang, Xiao-Ying Zhou, Wei-Ling Tang, Yi Sun, Guang-Ying Lu, Guang-Xiu Lu

ABSTRACT

How to maintain the genetic integrity of cultured human embryonic stem (hES) cells is raising crucial concerns for future clinical use in regenerative medicine. Mitomycin C(MMC), a DNA damage agent, is widely used for preparation of feeder cells in many laboratories. However, to what extent MMC affects the karyotypic stability of hES cells is not clear. Here, we measured residual MMC using High Performance Liquid Chromatography-Mass Spectrometry/Mass Spectrometry following each step of feeder preparation and found that 2.26 ± 0.77 and 3.50 ± 0.92 ng/ml remained in mouse feeder cells and human feeder cells, respectively. In addition, different amounts of MMC caused different chromosomal aberrations in hES cells. In particular, one abnormality, dup(1)(p32p36), was the same identical to one we previously reported in another hES cell line. Using Affymetrix SNP 6.0 arrays, the copy number variation changes of the hES cells maintained on MMC-inactivated feeders (MMC-feeder) were significantly more than those cultured on γ-inactivated feeder (IR-feeder) cells. Furthermore, DNA damage response (DDR) genes were down-regulated during long-term culture in the MMC-containing system, leading to DDR defect and shortened telomeres of hES cells, a sign of genomic instability. Therefore, MMC-feeder and MMC-induced genomic variation present an important safety problem that would limit such hES from being applied for future clinic use and drug screening. More... »

PAGES

33-45

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00204-014-1250-6

    DOI

    http://dx.doi.org/10.1007/s00204-014-1250-6

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1015549674

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/24838295


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