NANOG Is a Direct Target of TGFβ/Activin-Mediated SMAD Signaling in Human ESCs

Xu, Ren‐He; Sampsell-Barron, Tori L.; Gu, Feng; Root, Sierra H.; Peck, Ruthann M.; Pan, Guangjin; Yu, Junying; Antosiewicz‐Bourget, Jessica; Tian, Shulan; Stewart, Ron; Thomson, James A.

doi:10.1016/j.stem.2008.07.001

Cited by 457 publications

(420 citation statements)

References 42 publications

Supporting

Mentioning

379

Contrasting

Unclassified

Order By: Relevance

“…These include soluble protein growth factors [e.g., FGF2, TGF-β, and autocrine/ paracrine cues (15)] that signal via a number of pathways to support cell survival and pluripotency factors [e.g., Nanog, Oct4, Sox2 (38)(39)(40)]. In addition, hPSCs engage in cell-matrix interactions (31), and cell-cell contacts via cadherins are particularly important for survival and pluripotency maintenance.…”

Section: Discussionmentioning

confidence: 99%

A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation

Lei

Schaffer

2013

Proc. Natl. Acad. Sci. U.S.A.

294

312

View full text Add to dashboard Cite

Human pluripotent stem cells (hPSCs), including human embryonic stem cells and induced pluripotent stem cells, are promising for numerous biomedical applications, such as cell replacement therapies, tissue and whole-organ engineering, and high-throughput pharmacology and toxicology screening. Each of these applications requires large numbers of cells of high quality; however, the scalable expansion and differentiation of hPSCs, especially for clinical utilization, remains a challenge. We report a simple, defined, efficient, scalable, and good manufacturing practice-compatible 3D culture system for hPSC expansion and differentiation. It employs a thermoresponsive hydrogel that combines easy manipulation and completely defined conditions, free of any human-or animal-derived factors, and entailing only recombinant protein factors. Under an optimized protocol, the 3D system enables long-term, serial expansion of multiple hPSCs lines with a high expansion rate (∼20-fold per 5-d passage, for a 10 72 -fold expansion over 280 d), yield (∼2.0 × 10 7 cells per mL of hydrogel), and purity (∼95% Oct4+), even with single-cell inoculation, all of which offer considerable advantages relative to current approaches. Moreover, the system enabled 3D directed differentiation of hPSCs into multiple lineages, including dopaminergic neuron progenitors with a yield of ∼8 × 10 7 dopaminergic progenitors per mL of hydrogel and ∼80-fold expansion by the end of a 15-d derivation. This versatile system may be useful at numerous scales, from basic biological investigation to clinical development.H uman pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) (1) and induced pluripotent stem cells (iPSCs) (2), have the capacities for indefinite in vitro expansion and differentiation into all cell types within adults (3). They therefore represent highly promising cell sources for numerous biomedical applications, such as cell replacement therapies (4, 5), tissue and organ engineering (6), and pharmacology and toxicology screens (7,8). However, these applications require large numbers of cells of high quality (4, 6-8). For instance, ∼10 5 surviving dopaminergic (DA) neurons, ∼10 9 cardiomyocytes, or ∼10 9 beta cells are likely required to treat a patient with Parkinson disease (PD), myocardial infarction (MI), or type I diabetes, respectively (9). Additionally, far more cells are needed initially because both in vitro cell culture yields and subsequent in vivo survival of transplanted cells are typically very low. As examples of the latter, only ∼6% of transplanted dopaminergic neurons or ∼1% of injected cardiomyocytes reportedly survive in rodent models several months after transplantation (10, 11). Furthermore, there are large patient populations with degenerative diseases or organ failure (9), including over 1 million people with PD, 1-2.5 million with type I diabetes, and ∼8 million with MI in the United States alone (12). Large numbers of cells are also necessary for applications such as tissue engineering, where for ex...

show abstract

Section: Discussionmentioning

confidence: 99%

A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation

Lei

Schaffer

2013

Proc. Natl. Acad. Sci. U.S.A.

294

312

View full text Add to dashboard Cite

show abstract

“…Unlike the ambiguity surrounding FGF2 signalling, Smad2/3, the downstream effectors of Activin/Nodal signaling are previously known to directly bind and regulate the expression of NANOG [97]. Recent ChIPseq in hESCs also found the binding of SMAD2/3 at OCT4, TERT, MYC and DPPA4 genes, with SMAD2/3 sharing approximately one-third overlap with NANOG genomic targets [98].…”

Section: Signalling In Hescsmentioning

confidence: 99%

The transcriptional regulation of pluripotency

Yeo

2012

Cell Res

117

View full text Add to dashboard Cite

Abbreviations: 2C (2-cell); 2i (two inhibitor); BMP4 (bone morphogenic protein 4); ChIP (chromatin immunoprecipitation); ChIP-seq (chromatin immunoprecipitation and sequencing); ESCs (embryonic stem cells); EpiSCs (Epiblast-derived stem cells); Fgf4 (fibroblast growth factor 4); hESCs (human embryonic stem cells); ICM (inner cell mass); iPSCs (induced pluripotent stem cells); LIF (leukemia inhibitory factor); lincRNA (large intergenic non-coding RNA); mESCs (mouse embryonic stem cells); miRNA (micro RNA); ncRNA (non-coding RNA); PcG (Polycomb group proteins); PGCs (primodial germ cells); POU (Pit-Oct-Unc); RNAi (RNA interference); RNA-seq (RNA sequencing); SCF (stem cell factor); siRNA (short interfering RNA); XaXa (double X-chromosome active); XaXi (single X-chromosome inactivated)The defining features of embryonic stem cells (ESCs) are their self-renewing and pluripotent capacities. Indeed, the ability to give rise into all cell types within the organism not only allows ESCs to function as an ideal in vitro tool to study embryonic development, but also offers great therapeutic potential within the field of regenerative medicine. However, it is also this same remarkable developmental plasticity that makes the efficient control of ESC differentiation into the desired cell type very difficult. Therefore, in order to harness ESCs for clinical applications, a detailed understanding of the molecular and cellular mechanisms controlling ESC pluripotency and lineage commitment is necessary. In this respect, through a variety of transcriptomic approaches, ESC pluripotency has been found to be regulated by a system of ESC-associated transcription factors; and the external signalling environment also acts as a key factor in modulating the ESC transcriptome. Here in this review, we summarize our current understanding of the transcriptional regulatory network in ESCs, discuss how the control of various signalling pathways could influence pluripotency, and provide a future outlook of ESC research.

show abstract

“…Although several defined media have optimized the growth and maintenance condition for hESCs without the requirement of a feeder layer, these media still rely on bFGF supplementation. FGF signaling plays a critical role in the survival, self-renewal, proliferation, and maintenance of the undifferentiated hESC state in vitro (8)(9)(10)(11)(12)(13)(14).…”

mentioning

confidence: 99%

Overexpression of BCL2 enhances survival of human embryonic stem cells during stress and obviates the requirement for serum factors

Ardehali

Inlay

Ali

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The promise of pluripotent stem cells as a research and therapeutic tool is partly undermined by the technical challenges of generating and maintaining these cells in culture. Human embryonic stem cells (hESCs) are exquisitely sensitive to culture conditions, and require constant signaling by growth factors and cell-cell and cellmatrix interactions to prevent apoptosis, senescence, and differentiation. Previous work from our laboratory demonstrated that overexpression of the prosurvival gene BCL2 in mouse embryonic stem cells overrode the requirement of serum factors and feeder cells to maintain mESCs in culture. To determine whether this prosurvival gene could similarly protect hESCs, we generated hESC lines that constitutively or inducibly express BCL2. We find that BCL2 overexpression significantly decreases dissociation-induced apoptosis, resulting in enhanced colony formation from sorted single cells, and enhanced embryoid body formation. In addition, BCL2-hESCs exhibit normal growth in the absence of serum, but require basic fibroblast growth factor to remain undifferentiated. Furthermore, they maintain their pluripotency markers, form teratomas in vivo, and differentiate into all three germ layers. Our data suggest that the BCL2 signaling pathway plays an important role in inhibiting hESC apoptosis, such that its overexpression in hESCs offers both a survival benefit in conditions of stress by resisting apoptosis and obviates the requirement for serum or a feeder layer for maintenance.embryonic stem cell survival | viability

show abstract

NANOG Is a Direct Target of TGFβ/Activin-Mediated SMAD Signaling in Human ESCs

Cited by 457 publications

References 42 publications

A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation

A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation

The transcriptional regulation of pluripotency

Overexpression of BCL2 enhances survival of human embryonic stem cells during stress and obviates the requirement for serum factors

Contact Info

Product

Resources

About