Induced Neuronal Cells: How to Make and Define a Neuron

Yang, Nan; Ng, Yi Han; Pang, Zhiping P.; Südhof, Thomas C.; Wernig, Marius

doi:10.1016/j.stem.2011.11.015

Cited by 161 publications

(161 citation statements)

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“…To this end, it is conceivable that further optimization of the culturing conditions will lead to the generation of fully mature neurons. However, our data are perfectly in line with previous works describing the derivation of human iNCs (30,34,35), and support the idea that in general human cells are less plastic and require longer maturation compared with mouse cells (36,37).…”

Section: Discussionsupporting

confidence: 92%

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Giorgetti

Marchetto

et al. 2012

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

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The finding that certain somatic cells can be directly converted into cells of other lineages by the delivery of specific sets of transcription factors paves the way to novel therapeutic applications. Here we show that human cord blood (CB) CD133 + cells lose their hematopoietic signature and are converted into CB-induced neuronal-like cells (CB-iNCs) by the ectopic expression of the transcription factor Sox2, a process that is further augmented by the combination of Sox2 and c-Myc. Gene-expression analysis, immunophenotyping, and electrophysiological analysis show that CBiNCs acquire a distinct neuronal phenotype characterized by the expression of multiple neuronal markers. CB-iNCs show the ability to fire action potentials after in vitro maturation as well as after in vivo transplantation into the mouse hippocampus. This system highlights the potential of CB cells and offers an alternative means to the study of cellular plasticity, possibly in the context of drug screening research and of future cell-replacement therapies.neurons | reprogramming | stem cells T he fate of adult somatic cells is not fixed rigidly and can be reprogrammed by experimental manipulation. The generation of induced pluripotent stem cells (iPSCs) represents the most dramatic evidence that the epigenome of somatic cells are remarkably plastic (1). Recently, it has been reported that fibroblasts can be converted by ectopic expression of defined transcription factors into postmitotic neurons (2-9), neural progenitors (10, 11), and self-renewing neural stem cells (NSCs) (12, 13). However, most reported methods rely on the use of multiple transcription factors and the use of fibroblasts as donor cells.Here we investigate if cord blood (CB) stem cells can be induced to acquire a neuronal phenotype by using only one transcription factor. It has been previously shown that stem cell populations are more amenable to reprogramming than other somatic cells, probably as a result of their preexisting epigenetic state (14,15). Moreover, because of their biological characteristics and availability, CB cells as a donor cell type could offer clear logistic advantages vs. other adult somatic cell types (16,17). These cells can be collected without any risk for the donor and are young cells carrying minimal somatic mutations.Strikingly, we show that forced expression of Sox2 is sufficient to convert CB CD133 + cells into induced neuronal progenitor (NP)-like cells, a conversion process that is augmented by the coexpression of c-Myc. Sox2 is highly expressed in adult NSCs, is one of the earliest functional markers of neuroectodermal specification in the embryo, and plays a key role in the neural lineage specification (18)(19)(20). We show that Sox2-transduced CB cells acquire a distinct neuronal morphology and express multiple neuronal markers in vitro, and that they can be expanded for as many as 25 passages when cultured in permissive condition culture. Moreover, we show that CB-induced neuronal-like cells (CB-iNCs) are able to differentiate in vitro and ...

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Section: Discussionsupporting

confidence: 92%

Cord blood-derived neuronal cells by ectopic expression of Sox2 and c-Myc

Giorgetti

Marchetto

et al. 2012

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

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“…Moreover, efficient protocols now are available to guide the differentiation of iPS cells and embryonic stem (ES) cells into different types of neurons that then can be used for disease modeling (4-7). As a further development, we and others recently described direct conversion of nonneural cells into induced neuronal (iN) cells, which accelerates the generation of neurons from nonneuronal cells and renders this process more uniform and less variable (8)(9)(10)(11)(12)(13)(14)(15)(16). The iN cells behave like functional neurons in that they display neuron-like morphologies, express neuronal marker genes, fire action potentials (APs), and, at least in some instances, are not only the postsynaptic recipients of input synapses but also form presynaptic output synapses onto each other or onto cocultured primary neurons.…”

mentioning

confidence: 99%

Neurons generated by direct conversion of fibroblasts reproduce synaptic phenotype caused by autism-associated neuroligin-3 mutation

Chanda

Marro

Wernig

et al. 2013

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

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Recent studies suggest that induced neuronal (iN) cells that are directly transdifferentiated from nonneuronal cells provide a powerful opportunity to examine neuropsychiatric diseases. However, the validity of using this approach to examine disease-specific changes has not been demonstrated. Here, we analyze the phenotypes of iN cells that were derived from murine embryonic fibroblasts cultured from littermate wild-type and mutant mice carrying the autism-associated R704C substitution in neuroligin-3. We show that neuroligin-3 R704C-mutant iN cells exhibit a large and selective decrease in AMPA-type glutamate receptor-mediated synaptic transmission without changes in NMDA-type glutamate receptor-or in GABA A receptor-mediated synaptic transmission. Thus, the synaptic phenotype observed in R704C-mutant iN cells replicates the previously observed phenotype of R704C-mutant neurons. Our data show that the effect of the R704C mutation is applicable even to neurons transdifferentiated from fibroblasts and constitute a proof-of-concept demonstration that iN cells can be used for cellular disease modeling. induced pluripotent stem (iPS) cells now can be generated from nearly every somatic cell type (1-3). Moreover, efficient protocols now are available to guide the differentiation of iPS cells and embryonic stem (ES) cells into different types of neurons that then can be used for disease modeling (4-7). As a further development, we and others recently described direct conversion of nonneural cells into induced neuronal (iN) cells, which accelerates the generation of neurons from nonneuronal cells and renders this process more uniform and less variable (8-16). The iN cells behave like functional neurons in that they display neuron-like morphologies, express neuronal marker genes, fire action potentials (APs), and, at least in some instances, are not only the postsynaptic recipients of input synapses but also form presynaptic output synapses onto each other or onto cocultured primary neurons.The in vitro generation of neurons from nonneuronal cells is of great interest because of its potential for studying neural development and for producing neurons for regenerative medicine. A possibly even more important application may be the use of neurons transdifferentiated from nonneuronal cells for examining disease mechanisms, because transdifferentiated neurons can be obtained from living patients. Indeed, a large number of studies reported a multitude of disease-related changes in neurons differentiated from patient-derived iPS cells (17)(18)(19)(20), and even iN cells have been used for this purpose (13). However, to date no study has tested whether the neurons directly reprogrammed from fibroblasts, ES cells, or other nonneural cells actually can recapitulate a phenotype observed in primary neurons from the same organism.To address this important question, we asked whether iN cells derived from a mutant mouse with a defined synaptic phenotype can recapitulate the effect. We focused on a previously characterized autism-assoc...

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“…Most iNSCs established by reported methods are expandable, which provides the possibility of generating a large amount of cells at the neural progenitor stage for subsequent differentiation or manipulation. In contrast, iNs directly converted from fibroblasts are terminally committed neurons that are not expandable [9]. On the other hand, the in vitro cultured patient iNSCs can be used to study the specific phenotypes of disease-affected NSCs, which are impossible to obtain from patients directly.…”

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confidence: 99%