Abstract
Telomerase and telomeres are important for indefinite replication of stem cells. Recently, telomeres of somatic cells were found to be reprogrammed to elongate in induced pluripotent stem cells (iPSCs). The role of telomeres in developmental pluripotency in vivo of embryonic stem cells (ESCs) or iPSCs, however, has not been directly addressed. We show that ESCs with long telomeres exhibit authentic developmental pluripotency, as evidenced by generation of complete ESC pups as well as germline-competent chimeras, the most stringent tests available in rodents. ESCs with short telomeres show reduced teratoma formation and chimera production, and fail to generate complete ESC pups. Telomere lengths are highly correlated (r > 0.8) with the developmental pluripotency of ESCs. Short telomeres decrease the proliferative rate or capacity of ESCs, alter the expression of genes related to telomere epigenetics, down-regulate genes important for embryogenesis and disrupt germ cell differentiation. Moreover, iPSCs with longer telomeres generate chimeras with higher efficiency than those with short telomeres. Our data show that functional telomeres are essential for the developmental pluripotency of ESCs/iPSCs and suggest that telomere length may provide a valuable marker to evaluate stem cell pluripotency, particularly when the stringent tests are not feasible.
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Acknowledgements
This work was supported by MOST National Basic Research Program of China (973 project) (2009CB941000), James and Esther King Biomedical Research Program, and China Scholarship Council and NSFC (31000611) (JH).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Experimental design for ESC derivation and pluripotency test in vitro and in vivo. (PDF 29 kb)
Supplementary information, Figure S2
Isolation and genotyping of ESC lines. (PDF 172 kb)
Supplementary information, Figure S3
Telomere length of ESCs shown as relative telomere fluorescence intensity estimated by Q-FISH. (PDF 49 kb)
Supplementary information, Figure S4
Histological analysis of teratomas derived from WT, Het. (PDF 250 kb)
Supplementary information, Figure S5
Promoter methylation (A) and protein expression (B) of Nanog and Oct4. (PDF 107 kb)
Supplementary information, Figure S6
Germ cell differentiation in vitro. (PDF 28 kb)
Supplementary information, Figure S7
Characterization of ESCs from WT (Terc+/+), Het (Terc+/−), G1, G3 and G4 Terc−/− at passage 20. (PDF 79 kb)
Supplementary information, Table S1
Efficiency in derivation of ES cell lines from blastocysts (PDF 68 kb)
Supplementary information, Table S2
Karyotyping of ES cell lines (PDF 7 kb)
Supplementary information, Table S3
Original data of chimeras and ESC-mice produced from injection of ES cells into 8-cell recipient CD1 embryos (PDF 56 kb)
Supplementary information, Table S4
Telomere loss and chromosome fusion from telomerase deficient mouse ES cells at early passage (P6) (PDF 7 kb)
Supplementary information, Table S8
Pathway analysis of differentially expressed genes in G3/G4 Terc−/− versus WT or G1 Terc−/− ESCs (PDF 70 kb)
Supplementary information, Table S9
Primers for real-time PCR analysis (PDF 9 kb)
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Huang, J., Wang, F., Okuka, M. et al. Association of telomere length with authentic pluripotency of ES/iPS cells. Cell Res 21, 779–792 (2011). https://doi.org/10.1038/cr.2011.16
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DOI: https://doi.org/10.1038/cr.2011.16
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