Abstract
Induced pluripotent stem (iPS) cells have greatly provoked people’s interest due to their enormous potential of clinical applications. Increasing care is taken with the genetic safety of iPS cells. However, up to now, the chromosomal integrity of murine iPS (miPS) cells has been largely unknown. We have observed recurrent trisomy and/or Robertsonian translocation (Rb) of chromosome 14 in six out of nine independent miPS cell lines from three laboratories by G-banding, fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY) analyses, while all the miPS cell lines were derived from mouse embryonic fibroblasts (MEFs) or neural precursor cells (NPCs) with a normal karyotype. The miPS cells with trisomy and/or Rb of chromosome 14 showed growth advantage over the miPS cells with a normal karyotype. We found a significantly higher frequency of Rbs in the miPS cell lines induced with c-Myc than those without c-Myc. Our findings demonstrate that miPS cell lines have the propensity for chromosomal aberrations and there is an obvious correlation between the extent of chromosomal aberrations in miPS cells and the transcriptional factors used for their reprogramming. Therefore, our study raises awareness of the need for improvements of the induction conditions of miPS cells in order to avoid the chromosomal aberrations and ensure future safe applications.
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Abbreviations
- ES:
-
Embryonic stem
- iPS:
-
Induced pluripotent stem
- MEFs:
-
Mouse embryonic fibroblasts
- NPCs:
-
Neural precursor cells
- Oct-4:
-
Octamer-binding transcription factor 4
- Sox2:
-
SRY (sex determining region Y)-box 2
- Klf4:
-
Kruppel-like factor 4
- c-Myc:
-
Cellular myelocytomatosis oncogene
- Rb:
-
Robertsonian translocation
- FISH:
-
Fluorescence in situ hybridization
- SKY:
-
Spectral karyotyping
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Acknowledgements
This work was supported by the National Natural Science Foundation of China (30711120571, 30725013 and 30671168), the National Basic Research Program (2007CB947401) of China (973) and the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy) at Hannover Medical School. We would like to thank Professor Julang Li (University of Guelph) for helpful discussions, Professor Guoliang Xu, Ying Jin and Jinsong Li (Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences) for providing mouse-induced pluripotent stem cell lines, and Professor Shan Zha (Princeton University) for providing BAC clones.
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Responsible Editor: Fengtang Yang
Qian Chen and Xiaoyun Shi equally contributed to this study.
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Fig. S1
Pluripotency of murine iPS cells. a Examples of iPS cells that maintain characteristic mESC-colony morphology and express pluripotency markers Oct4, Nanog, Sox2 and SSEA1. b Hematoxylin and eosin staining of teratoma derived from iPS3 (upper lane) and iPS6 cells (lower lane); scale bar is 50 μm (JPEG 47 kb)
Fig. S2
The quality of SKY is evaluated by using the classified images. Representative SKY images of miPS cells showing a 41, XY, +14 (iPS2), and b 40, XY, +14, Rb(14.14) (iPS5). Each chromosome is shown in spectral image (left), inverted DAPI (middle), and after classification (right) (JPEG 38 kb)
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Chen, Q., Shi, X., Rudolph, C. et al. Recurrent trisomy and Robertsonian translocation of chromosome 14 in murine iPS cell lines. Chromosome Res 19, 857–868 (2011). https://doi.org/10.1007/s10577-011-9239-y
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DOI: https://doi.org/10.1007/s10577-011-9239-y