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Alternative routes to induce naive pluripotency in human embryonic stem cells

(2015) STEM CELLS. 33(9). p.2686-2698
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Abstract
Human embryonic stem cells (hESCs) closely resemble mouse epiblast stem cells exhibiting primed pluripotency unlike mouse ESCs (mESCs), which acquire a naive pluripotent state. Efforts have been made to trigger naive pluripotency in hESCs for subsequent unbiased lineage-specific differentiation, a common conundrum faced by primed pluripotent hESCs due to heterogeneity in gene expression existing within and between hESC lines. This required either ectopic expression of naive genes such as NANOG and KLF2 or inclusion of multiple pluripotency-associated factors. We report here a novel combination of small molecules and growth factors in culture medium (2i/LIF/basic fibroblast growth factor1Ascorbic Acid1Forskolin) facilitating rapid induction of transgene-free naive pluripotency in hESCs, as well as in mESCs, which has not been shown earlier. The converted naive hESCs survived long-term single-cell passaging, maintained a normal karyotype, upregulated naive pluripotency genes, and exhibited dependence on signaling pathways similar to naive mESCs. Moreover, they undergo global DNA demethylation and show a distinctive long noncoding RNA profile. We propose that in our medium, the FGF signaling pathway via PI3K/AKT/mTORC induced the conversion of primed hESCs toward naive pluripotency. Collectively, we demonstrate an alternate route to capture naive pluripotency in hESCs that is fast, reproducible, supports naive mESC derivation, and allows efficient differentiation.
Keywords
Small molecules, Signaling, GROUND-STATE PLURIPOTENCY, KINASE-ACTIVITY, SMALL MOLECULES, SELF-RENEWAL, EPIBLAST, DIFFERENTIATION, DERIVATION, MAINTENANCE, MICROARRAY, CONVERSION, Human embryonic stem cells, Naive, Pluripotency

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Chicago
Duggal, Galbha, Sharat Warrier, Sabitri Ghimire, Dorien Broekaert, Margot Van der Jeught, Sylvie Lierman, Tom Deroo, et al. 2015. “Alternative Routes to Induce Naive Pluripotency in Human Embryonic Stem Cells.” Stem Cells 33 (9): 2686–2698.
APA
Duggal, G., Warrier, S., Ghimire, S., Broekaert, D., Van der Jeught, M., Lierman, S., Deroo, T., et al. (2015). Alternative routes to induce naive pluripotency in human embryonic stem cells. STEM CELLS, 33(9), 2686–2698.
Vancouver
1.
Duggal G, Warrier S, Ghimire S, Broekaert D, Van der Jeught M, Lierman S, et al. Alternative routes to induce naive pluripotency in human embryonic stem cells. STEM CELLS. 2015;33(9):2686–98.
MLA
Duggal, Galbha, Sharat Warrier, Sabitri Ghimire, et al. “Alternative Routes to Induce Naive Pluripotency in Human Embryonic Stem Cells.” STEM CELLS 33.9 (2015): 2686–2698. Print.
@article{7021797,
  abstract     = {Human embryonic stem cells (hESCs) closely resemble mouse epiblast stem cells exhibiting primed pluripotency unlike mouse ESCs (mESCs), which acquire a naive pluripotent state. Efforts have been made to trigger naive pluripotency in hESCs for subsequent unbiased lineage-specific differentiation, a common conundrum faced by primed pluripotent hESCs due to heterogeneity in gene expression existing within and between hESC lines. This required either ectopic expression of naive genes such as NANOG and KLF2 or inclusion of multiple pluripotency-associated factors. We report here a novel combination of small molecules and growth factors in culture medium (2i/LIF/basic fibroblast growth factor1Ascorbic Acid1Forskolin) facilitating rapid induction of transgene-free naive pluripotency in hESCs, as well as in mESCs, which has not been shown earlier. The converted naive hESCs survived long-term single-cell passaging, maintained a normal karyotype, upregulated naive pluripotency genes, and exhibited dependence on signaling pathways similar to naive mESCs. Moreover, they undergo global DNA demethylation and show a distinctive long noncoding RNA profile. We propose that in our medium, the FGF signaling pathway via PI3K/AKT/mTORC induced the conversion of primed hESCs toward naive pluripotency. Collectively, we demonstrate an alternate route to capture naive pluripotency in hESCs that is fast, reproducible, supports naive mESC derivation, and allows efficient differentiation.},
  author       = {Duggal, Galbha and Warrier, Sharat and Ghimire, Sabitri and Broekaert, Dorien and Van der Jeught, Margot and Lierman, Sylvie and Deroo, Tom and Peelman, Luc and Van Soom, Ann and Cornelissen, Maria and Menten, Bj{\"o}rn and Mestdagh, Pieter and Vandesompele, Jo and Roost, Matthias and Slieker, Roderick C and Heijmans, Bastiaan T and Deforce, Dieter and De Sutter, Petra and Chuva de Sousa Lopes, Susana Marina and Heindryckx, Bj{\"o}rn},
  issn         = {1066-5099},
  journal      = {STEM CELLS},
  keyword      = {Small molecules,Signaling,GROUND-STATE PLURIPOTENCY,KINASE-ACTIVITY,SMALL MOLECULES,SELF-RENEWAL,EPIBLAST,DIFFERENTIATION,DERIVATION,MAINTENANCE,MICROARRAY,CONVERSION,Human embryonic stem cells,Naive,Pluripotency},
  language     = {eng},
  number       = {9},
  pages        = {2686--2698},
  title        = {Alternative routes to induce naive pluripotency in human embryonic stem cells},
  url          = {http://dx.doi.org/10.1002/stem.2071},
  volume       = {33},
  year         = {2015},
}

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