Pluripotency Reprogramming
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Generation of eggs from mouse embryonic stem cells and induced pluripotent stem cells

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Nanog Heterogeneity: Tilting at Windmills?

Austin Smith's comments: Cell 2013

Importantly, there is currently no evidence that either fluctuating expression of pluripotency factors or state reversion occurs during epiblast progression and lineage commitment in vivo. Instead, the variation that many have observed may simply be a culture epiphenomenon: an attractive playbox for experimentalists and modellers, but with questionable relevance for the way in which pluripotent cells really make fate decisions.

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Klf4 Organizes Long-Range Chromosomal Interactions with the Oct4 Locus in Reprogramming and Pluripotency

Klf4 Organizes Long-Range Chromosomal Interactions with the Oct4 Locus in Reprogramming and Pluripotency | Pluripotency Reprogramming | Scoop.it
Summary

Epigenetic mechanisms underlying somatic reprogramming have been extensively studied, but little is known about the nuclear architecture of pluripotent stem cells (PSCs). Using circular chromosome conformation capture with high-throughput sequencing (4C-seq) and fluorescence in situ hybridization (FISH), we identified chromosomal regions that colocalize frequently with the Oct4 locus in PSCs. These PSC-specific long-range interactions are established prior to transcriptional activation of endogenous Oct4 during reprogramming to induced PSCs and are facilitated by Klf4-mediated recruitment of cohesin. Depletion of Klf4 leads to unloading of cohesin at the Oct4 enhancer and disrupts long-range interactions prior to loss of Oct4 transcription and subsequent PSC differentiation, suggesting a causative role for Klf4 in facilitating long-range interactions independent of its transcriptional activity. Taken together, our results delineate the basic nuclear organization at the Oct4 locus in PSCs and suggest a functional role for Klf4-mediated higher-order chromatin structure in maintaining and inducing pluripotency.

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TaqMan® hPSC Scorecard™ Panel and data analysis - Characterize stem cell lines relative to a standard

TaqMan® hPSC Scorecard™ Panel and data analysis - Characterize stem cell lines relative to a standard | Pluripotency Reprogramming | Scoop.it

Overview of protocol:

1 Test pluripotent, random differentiation (EB) or inductive differentiation samples

2 Extract and purify RNA

3 Generate cDNA

4 Run gene expression panel on QuantStudio™ 12K Flex, ViiA™ 7 or StepOnePlus™ Real Time PCR Systems

5 Upload .eds files to the hPSC Scorecard™ Analysis Software

6 Analyze results using the intuitive hPSC Scorecard™ Software Analysis Tool

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Stemgent - These stem cell scientists are successfully producing non-integrated iPS cell lines using mRNA reprogramming models.

Stemgent - These stem cell scientists are successfully producing non-integrated iPS cell lines using mRNA reprogramming models. | Pluripotency Reprogramming | Scoop.it
Stemgent has been working alongside some of the world's leading stem cell scientists in developing innovative technology and application solutions for the advancement of stem cell and iPS cell research.
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Cell Stem Cell - Browsing (Epi)genomes: A Guide to Data Resources and Epigenome Browsers for Stem Cell Researchers

Cell Stem Cell - Browsing (Epi)genomes: A Guide to Data Resources and Epigenome Browsers for Stem Cell Researchers | Pluripotency Reprogramming | Scoop.it
Karnik R and Meissner A, Cell 2013

This review is to provide the reader with information on where to find many of the data sets and how to utilize the various (epi)genome browsers for display and initial analysis. We provide selected examples to highlight key features and demonstrate the application of these browsers to stem cell biology.

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Cell Stem Cell - Single-Cell Analysis Reveals that Expression of Nanog Is Biallelic and Equally Variable as that of Other Pluripotency Factors in Mouse ESCs

Cell Stem Cell - Single-Cell Analysis Reveals that Expression of Nanog Is Biallelic and Equally Variable as that of Other Pluripotency Factors in Mouse ESCs | Pluripotency Reprogramming | Scoop.it
Highlights

 

Single-cell analysis reveals that Nanog is biallelically expressed in mESCs

Variation in Nanog expression is similar to that of other pluripotency markers

Heterozygous loss-of-function knockin reporters do not reflect Nanog expression

Genetic background can influence the range of Nanog expression

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Concise Review: Embryonic Stem Cells Versus Induced Pluripotent Stem Cells: The Game Is On - Puri - 2011 - STEM CELLS - Wiley Online Library

Concise Review: Embryonic Stem Cells Versus Induced Pluripotent Stem Cells: The Game Is On - Puri - 2011 - STEM CELLS - Wiley Online Library | Pluripotency Reprogramming | Scoop.it

Permanent cell lines of pluripotent ESCs and iPSCs and our increasing ability to direct them into any cell type for therapeutic potential holds enormous promise for future regenerative medicine. ESCs are considered to be the gold standard of pluripotency, while iPSCs offer the development of cells from any adult individual, which advances the possibility of curing devastating degenerative diseases using cell or tissue grafts with perfect histocompatibility match. This potential calls for efforts to characterize and compare the nature of these pluripotent cell types in great detail. Only such deep studies can give us sufficient insight into the potential, efficacy, and safety to reach a decision; which one will be more favorable for future clinical applications. At the current state of knowledge, we are not in a position to make such a decision. The game between ESCs and iPSCs is still on with no obvious indication of the winner.

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Stemgent - Interview With Jun Liu

Stemgent - Interview With Jun Liu | Pluripotency Reprogramming | Scoop.it
Jun Liu, PhD at Monash Institute of Medical Research Interview on mRNA Reprogramming Success.
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