Embryonic Stem Cell‐Derived Neural Progenitors Display Temporal Restriction to Neural Patterning

Bouhon, Isabelle Anne; Joannides, Alexis; Kato, Hidemasa; Chandran, Siddharthan; Allen, Nicholas Denby

doi:10.1634/stemcells.2006-0031

Cited by 52 publications

(51 citation statements)

References 42 publications

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“…The underlying problem is most probably related to the short time window following neural induction during which the cells can be directed towards specific neuronal lineages (Bouhon et al 2006); after this time, they become unresponsive to most developmental patterning cues.…”

Section: Neural Stem Cellsmentioning

confidence: 99%

“…Long-term expansion of ES cells seems to present less of a problem, since the cells maintain a pluripotent phenotype, thus allowing the production of large numbers of cells, and their differentiation potential seems to become restricted only once they are directed towards a neural phenotype (Bouhon et al 2006). However, the accumulation of chromosomal abnormalities in ES cells over continued passages has been reported (Draper et al 2004) and the efficiency with which different human stem cell lines generate different neural and neuronal phenotypes is known to differ (Iacovitti et al 2007;Park et al 2005).…”

Section: Maintenance Of Differentiated Cells Long-termmentioning

confidence: 99%

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Human stem cells for CNS repair

et al. 2007

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Although most peripheral tissues have at least a limited ability for self-repair, the central nervous system (CNS) has long been known to be relatively resistant to regeneration. Small numbers of stem cells have been found in the adult brain but do not appear to be able to affect any significant recovery following disease or insult. In the last few decades, the idea of being able to repair the brain by introducing new cells to repair damaged areas has become an accepted potential treatment for neurodegenerative diseases. This review focuses on the suitability of various human stem cell sources for such treatments of both slowly progressing conditions, such as Parkinson's disease, Huntington's disease and multiple sclerosis, and acute insult, such as stroke and spinal cord injury. Despite stem cell transplantation having now moved a step closer to the clinic with the first trials of autologous mesenchymal stem cells, the effects shown are moderate and are not yet at the stage of development that can fulfil the hopes that have been placed on stem cells as a means to replace degenerating cells in the CNS. Success will depend on careful investigation in experimental models to enable us to understand not just the practicalities of stem cell use, but also the underlying biological principles.

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Section: Neural Stem Cellsmentioning

confidence: 99%

Section: Maintenance Of Differentiated Cells Long-termmentioning

confidence: 99%

Human stem cells for CNS repair

et al. 2007

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“…In vitro differentiation of mESCs gives rise to the three primary germ layers -the endoderm, mesoderm and ectoderm -that characterise early stages of embryonic development (Leahy et al, 1999). This is then followed by the expression of stagespecific markers, associated with certain lineages, such as vimentin, nestin and ␤-tubulin III during neuronal differentiation (Rolletschek et al, 2001;Bouhon et al, 2006) and brachyury during cardiac differentiation Wilkinson et al, 1990). There is some initial evidence to suggest that POLG and TFAM are both expressed during early hESC differentiation, as is OCT4 (St John et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial DNA replication during differentiation of murine embryonic stem cells

Facucho-Oliveira

Alderson

Spikings

et al. 2007

Journal of Cell Science

259

261

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Oxidative phosphorylation (OXPHOS), the intracellular process that generates the majority of the ATP of a cell through the electron-transfer chain, is highly dependent on proteins encoded by the mitochondrial genome (mtDNA). MtDNA replication is regulated by the nuclear-encoded mitochondrial transcription factor A (TFAM) and the mitochondrial-specific DNA polymerase gamma, which consists of a catalytic (POLG) and an accessory (POLG2) subunit. Differentiation of pluripotent embryonic stem cells (ESCs) into specific cell types requires expansion of discrete populations of mitochondria and mtDNA replication to meet the specific metabolic requirements of the cell. We determined by real-time PCR that expression of pluripotent markers is reduced before the upregulation of Polg, Polg2 and Tfam in spontaneously differentiating R1 murine (m)ESCs, along with transient increases in mtDNA copy number. In D3 mESCs, the initial transient increase did not take place. However, precursors of neuronal and cardiomyocyte differentiation were positive for both POLG and TFAM. Similar-stage ESCs also showed active mtDNA replication, identified by 5-bromo-2′-deoxy-uridine labelling, as mtDNA copy number increased. Retinoic-acid-induced differentiation resulted in more consistent patterns of replication and upregulation of Polg, Polg2 and Tfam, whereas siRNA knockdown demonstrated that steady-state expression of POLG is essential for maintaining pluripotency.

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“…Neuronal differentiation experiments were performed as described previously (Bouhon et al 2006;Kohama et al 2012) with a minor modification. Briefly, after trypsinization of MB4 cultured on MEF plates, cell mixtures were transferred to new cell culture dishes twice and incubated for 10 min after the first transfer and 15 min after the second transfer in a CO 2 incubator to deplete MEF.…”

Section: Neuronal Differentiationmentioning

confidence: 99%

Comparison of two types of non-adherent plate for neuronal differentiation of mouse embryonic stem cells

2016

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In vitro differentiation systems of mouse embryonic stem cells (ESCs) are widely used as tools for studies of cell differentiation, organogenesis, and regenerative medicine. We have studied the regulation of neuron-specific imprinting genes, Ube3a and its antisense transcripts (Ube3a ATS), using in vitro neuronal differentiation of F1 hybrid ESCs. Each different non-adherent plate used for embryoid body (EB) formation during differentiation is associated with different costs; notably, plates coated with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer are more expensive than untreated polystyrene plates. Here, we assessed whether the polymercoated plates gave better results than the untreated plates. The first stage of differentation was performed in the MPC polymer-coated or untreated plates. The formed EBs were then passaged onto laminin-coated plates for further differentiation into neurons. Neither the neuron-specific imprinting status of Ube3a nor the expression levels of the neuron-specific markers Ube3a ATS and Mtap2 differed between neurons prepared on untreated plates and those prepared on MPC polymer-coated plates. These results suggest that the two non-adherent plates displayed almost the same characteristics for inducing neuronal differentiation of mouse ESCs and EB formation. Our study proved that untreated polystyrene plates are a costeffective choice for EB formation in in vitro differentiation systems of mouse ESCs.

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Embryonic Stem Cell‐Derived Neural Progenitors Display Temporal Restriction to Neural Patterning

Cited by 52 publications

References 42 publications

Human stem cells for CNS repair

Human stem cells for CNS repair

Mitochondrial DNA replication during differentiation of murine embryonic stem cells

Comparison of two types of non-adherent plate for neuronal differentiation of mouse embryonic stem cells

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