Viable mice produced from three-factor induced pluripotent stem (iPS) cells through tetraploid complementation

Kang, Lan; Wu, Tong; Yu, Tao; Yuan, Ye; He, Jing; Luo, Tong; Kou, Zhaohui; Gao, Shaorong

doi:10.1038/cr.2010.164

Cited by 32 publications

(32 citation statements)

References 11 publications

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“…These iPSC lines give rise to viable pups with efficiencies of 5-13%, which is comparable to ESCs 3,4,7 and higher than that reported for most other iPSC lines [8][9][10][11][12] . These reports show that direct reprogramming can produce fully pluripotent iPSCs that match ESCs in their developmental potential and efficiency of generating pups in TEC tests.…”

Section: Introductionmentioning

confidence: 80%

“…This allows us to identify lines with multiple copies of different lentiviruses while limiting toxicity to the MEFs and producing colonies without overcrowding the wells. It should be noted that multiple other protocols have been shown to produce iPSCs with full developmental potential, using multiple methods and donor cell sources suggesting that multiple paths to full pluripotency may exist 1,[8][9][10][11][12][13]15 . At present, however, no definitive biomarker of fully pluripotent iPSC has been identified and therefore the TEC assay remains the gold standard test of whether an iPSC line can generate all cell lineages in an organism.…”

Section: Discussionmentioning

confidence: 99%

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Generation of Mice Derived from Induced Pluripotent Stem Cells

Boland

Hazen

Nazor

et al. 2012

JoVE

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The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also produce a source of patient-matched cells for regenerative therapies. iPSCs may be generated using multiple protocols and derived from multiple cell sources. Once generated, iPSCs are tested using a variety of assays including immunostaining for pluripotency markers, generation of three germ layers in embryoid bodies and teratomas, comparisons of gene expression with embryonic stem cells (ESCs) and production of chimeric mice with or without germline contribution 2 . Importantly, iPSC lines that pass these tests still vary in their capacity to produce different differentiated cell types 2 . This has made it difficult to establish which iPSC derivation protocols, donor cell sources or selection methods are most useful for different applications.The most stringent test of whether a stem cell line has sufficient developmental potential to generate all tissues required for survival of an organism (termed full pluripotency) is tetraploid embryo complementation (TEC) [3][4][5] . Technically, TEC involves electrofusion of two-cell embryos to generate tetraploid (4n) one-cell embryos that can be cultured in vitro to the blastocyst stage 6 . Diploid (2n) pluripotent stem cells (e.g. ESCs or iPSCs) are then injected into the blastocoel cavity of the tetraploid blastocyst and transferred to a recipient female for gestation (see Figure 1). The tetraploid component of the complemented embryo contributes almost exclusively to the extraembryonic tissues (placenta, yolk sac), whereas the diploid cells constitute the embryo proper, resulting in a fetus derived entirely from the injected stem cell line.Recently, we reported the derivation of iPSC lines that reproducibly generate adult mice via TEC 1 . These iPSC lines give rise to viable pups with efficiencies of 5-13%, which is comparable to ESCs 3,4,7 and higher than that reported for most other iPSC lines [8][9][10][11][12] . These reports show that direct reprogramming can produce fully pluripotent iPSCs that match ESCs in their developmental potential and efficiency of generating pups in TEC tests. At present, it is not clear what distinguishes between fully pluripotent iPSCs and less potent lines [13][14][15] . Nor is it clear which reprogramming methods will produce these lines with the highest efficiency. Here we describe one method that produces fully pluripotent iPSCs and "all-iPSC" mice, which may be helpful for investigators wishing to compare the pluripotency of iPSC lines or establish the equivalence of different reprogramming methods. Video LinkThe video component of this article can be found at https://www.jove.com/video/4003/ ProtocolThis method was used in the research reported in Boland et al. Nature. 461, 91-96 (2009 Preparation of LentivirusThis protocol employs doxycycline-inducible lentiviral shuttle vectors that encode for Oct4, Sox2, Klf4, and c-Myc under control of a tetO response element. Tran...

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Section: Introductionmentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

Generation of Mice Derived from Induced Pluripotent Stem Cells

Boland

Hazen

Nazor

et al. 2012

JoVE

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“…However, almost all currently available high-quality iPSC lines, as determined by germline transmission or tetraploid blastocyst complementation, are male [3,[14][15][16][17][18][19][20]. Most of these iPSCs were generated from gender-mixed somatic cells.…”

Section: Introductionmentioning

confidence: 98%

“…In addition, some iPSCs can attain true pluripotency and support the development of all-iPSC mice using tetraploid complementation, the most stringent assay for developmental potential. This generation of iPSCs has revolutionized the field of regenerative medicine and also provides a unique platform to study genetic diseases in vitro, because they are technically simple to generate and are ethically non-controversial.However, almost all currently available high-quality iPSC lines, as determined by germline transmission or tetraploid blastocyst complementation, are male [3,[14][15][16][17][18][19][20]. Most of these iPSCs were generated from gender-mixed somatic cells.…”

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confidence: 98%

Generation of Fully Pluripotent Female Murine-Induced Pluripotent Stem Cells1

Gao

Cui

et al. 2015

Biology of Reproduction

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The high quality of induced pluripotent stem cells (iPSCs) has been determined to be high-grade chimeras that are competent for germline transmission, and viable mice can be generated through tetraploid complementation. Most of the high-quality iPSCs described to date have been male. Female iPSCs, especially fully pluripotent female iPSCs, are also essential for clinical applications and scientific research. Here, we show, for the first time, that a gender-mixed induction strategy could lead to a skewed sex ratio of iPSCs. After reprogramming, 50%, 70%, and 90% female initiating mouse embryonic fibroblasts at different male ratios resulted in 14.1 ± 6.8% (P < 0.05), 31.8 ± 5.4% (P < 0.05), and 80.1 ± 2.8% (P < 0.05) female iPSCs, respectively. Furthermore, these female iPSCs had pluripotent properties typical of embryonic stem cells. Importantly, these fully pluripotent female iPSCs could generate viable mice by tetraploid complementation. These findings indicate that high-quality female iPSCs could be derived effectively, and suggest that clinical application of female iPSCs is feasible.

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“…Although the pluripotency of most iPS cell lines that were tested correlated well with the expression status of these genes, some exceptions existed, in particular Oct4, Sox2 and Klf4 used to derive iPS cell lines [26]. Moreover, the fact that Gtl2 knockout mice are viable challenges the importance of Gtl2 in determining the pluripotency of iPS cells [28].…”

Section: Kang and Gaomentioning

confidence: 99%

Pluripotency of Induced Pluripotent Stem Cells

Kang

Gao

2012

Journal of Animal Science and Biotechnology

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Recent studies have demonstrated that differentiated somatic cells from various mammalian species can be reprogrammed into induced pluripotent stem (iPS) cells by the ectopic expression of four transcription factors that are highly expressed in embryonic stem (ES) cells. The generation of patient-specific iPS cells directly from somatic cells without using oocytes or embryos holds great promise for curing numerous diseases that are currently unresponsive to traditional clinical approaches. However, some recent studies have argued that various iPS cell lines may still retain certain epigenetic memories that are inherited from the somatic cells. Such observations have raised concerns regarding the safety and efficacy of using iPS cell derivatives for clinical applications. Recently, our study demonstrated full pluripotency of mouse iPS cells by tetraploid complementation, indicating that it is possible to obtain fully reprogrammed iPS cells directly from differentiated somatic cells. Therefore, we propose in this review that further comprehensive studies of both mouse and human iPS cells are required so that additional information will be available for evaluating the quality of human iPS cells.

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Viable mice produced from three-factor induced pluripotent stem (iPS) cells through tetraploid complementation

Cited by 32 publications

References 11 publications

Generation of Mice Derived from Induced Pluripotent Stem Cells

Generation of Mice Derived from Induced Pluripotent Stem Cells

Generation of Fully Pluripotent Female Murine-Induced Pluripotent Stem Cells1

Pluripotency of Induced Pluripotent Stem Cells

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