Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells

Xu, Ren‐He; Peck, Ruthann M.; Li, Dong S.; Feng, Xuezhu; Ludwig, Tenneille E.; Thomson, James A.

doi:10.1038/nmeth744

Cited by 906 publications

(758 citation statements)

References 28 publications

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“…Further comparing Noggin-treated cells with nontreated cells revealed that granulocyte colony-stimulating factor (G-CSF) promotes the proliferation of developing cardiomyocytes derived from mESCs [30]. Noggin sustains the undifferentiated proliferation of hESCs [31], and BMP4 is required for mESC self-renewal [32]. These different self-renewal mechanisms may be the cause of the differences observed in the cardiac differentiation studies of human and mouse embryonic stem cells.…”

Section: Discussionmentioning

confidence: 99%

Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals

et al. 2010

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Although myocyte cell transplantation studies have suggested a promising therapeutic potential for myocardial infarction, a major obstacle to the development of clinical therapies for myocardial repair is the difficulties associated with obtaining relatively homogeneous ventricular myocytes for transplantation. Human embryonic stem cells (hESCs) are a promising source of cardiomyocytes. Here we report that retinoid signaling regulates the fate specification of atrial versus ventricular myocytes during cardiac differentiation of hESCs. We found that both Noggin and the panretinoic acid receptor antagonist BMS-189453 (RAi) significantly increased the cardiac differentiation efficiency of hESCs. To investigate retinoid functions, we compared Noggin+RAi-treated cultures with Noggin+RA-treated cultures. Our results showed that the expression levels of the ventricular-specific gene IRX-4 were radically elevated in Noggin+RAi-treated cultures. MLC-2V, another ventricular-specific marker, was expressed in the majority of the cardiomyocytes in Noggin+RAi-treated cultures, but not in the cardiomyocytes of Noggin+RA-treated cultures. Flow cytometry analysis and electrophysiological studies indicated that with 64.7 ± 0.88% (mean ± s.e.m) cardiac differentiation efficiency, 83% of the cardiomyocytes in Noggin+RAi-treated cultures had embryonic ventricular-like action potentials (APs). With 50.7 ± 1.76% cardiac differentiation efficiency, 94% of the cardiomyocytes in Noggin+RA-treated cultures had embryonic atrial-like APs. These results were further confirmed by imaging studies that assessed the patterns and properties of the Ca 2+ sparks of the cardiomyocytes from the two cultures. These findings demonstrate that retinoid signaling specifies the atrial versus ventricular differentiation of hESCs. This study also shows that relatively homogeneous embryonic atrial-and ventricular-like myocyte populations can be efficiently derived from hESCs by specifically regulating Noggin and retinoid signals.

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Section: Discussionmentioning

confidence: 99%

Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals

et al. 2010

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“…Cells cultured in this manner were found to exhibit a normal karyotype and were able to form both EBs and teratomas. Xu, Peck, et al (2005) carried out a similar study in which they stated that, in unconditioned medium, hESCs are subjected to high levels of BMP signaling, which are reduced in MEF-CM. They found that a combination of the BMP antagonist, Noggin (500 ng/ml), and bFGF (40 ng/ml) was sufficient to sustain undifferentiated growth of several WiCell lines on Matrigel.…”

Section: Defined Mediummentioning

confidence: 99%

“…This role for FGF may involve activation of the PI-3-kinase/Akt pathway (Kim et al, 2005) although direct evidence for this has to be established. Finally, inhibition of the BMP signaling pathway, which is suggested to drive differentiation of hESCs, may also be important (Xu, Peck, et al, 2005). Investigation and manipulation of these pathways will be useful in the drive to feeder-free culture in defined culture environments.…”

Section: Analyzing the Mef Contributionmentioning

confidence: 99%

Toward xeno-free culture of human embryonic stem cells

Mallon

Park

Chen

et al. 2006

The International Journal of Biochemistry & Cell Biology

158

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The culture of human embryonic stem cells (hESCs) is limited, both technically and with respect to clinical potential, by the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. The concern over xenogeneic contaminants from the mouse feeder cells may restrict transplantation to humans and the variability in MEFs from batch-to-batch and laboratory-to-laboratory may contribute to some of the variability in experimental results. Finally, use of any feeder layer increases the work load and subsequently limits the large-scale culture of human ES cells. Thus, the development of feeder-free cultures will allow more reproducible culture conditions, facilitate scale-up and potentiate the clinical use of cells differentiated from hESC cultures. In this review, we describe various methods tested to culture cells in the absence of MEF feeder layers and other advances in eliminating xenogeneic products from the culture system.

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“…However, for human ESCs, other exogenous growth factors than LIF and BMP4 are required for their maintenance. For example, bFGF [81,82] , TGF-β/activin A [83,84] , Wnt [81] proteins and insulinlike growth factor [85] are reported to support the self-renewal of hESCs.…”

Section: Small Molecules Promoting Self-renewal Of Escs/ipscsmentioning

confidence: 99%

iPSCs and small molecules: a reciprocal effort towards better approaches for drug discovery

Zhang

Xie

2013

Acta Pharmacol Sin

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The revolutionary induced pluripotent stem cell (iPSC) technology provides a new path for cell replacement therapies and drug screening. Patient-specific iPSCs and subsequent differentiated cells manifesting disease phenotypes will finally position human disease pathology at the core of drug discovery. Cells used to test the toxic effects of drugs can also be generated from normal iPSCs and provide a much more accurate and cost-effective system than many animal models. Here, we highlight the recent progress in iPSC-based cell therapy, disease modeling and drug evaluations. In addition, we discuss the use of small molecule drugs to improve the generation of iPSCs and understand the reprogramming mechanism. It is foreseeable that the interplay between iPSC technology and small molecule compounds will push forward the applications of iPSC-based therapy and screening systems in the real world and eventually revolutionize the methods used to treat diseases.

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Basic FGF and suppression of BMP signaling sustain undifferentiated proliferation of human ES cells

Cited by 906 publications

References 28 publications

Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals

Direct differentiation of atrial and ventricular myocytes from human embryonic stem cells by alternating retinoid signals

Toward xeno-free culture of human embryonic stem cells

iPSCs and small molecules: a reciprocal effort towards better approaches for drug discovery

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