Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks

Shi, Yunfeng; Kirwan, Peter; Livesey, Frederick J.

doi:10.1038/nprot.2012.116

Cited by 807 publications

(898 citation statements)

References 24 publications

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“…In the past decade, several protocols to convert hiPSCs into neurons have been developed [2][3][4][5][6][7][8] . However, these protocols are still limited in many ways.…”

Section: Introductionmentioning

confidence: 99%

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

Frega

Gestel

Linda

et al. 2017

JoVE

115

189

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Neurons derived from human induced Pluripotent Stem Cells (hiPSCs) provide a promising new tool for studying neurological disorders. In the past decade, many protocols for differentiating hiPSCs into neurons have been developed. However, these protocols are often slow with high variability, low reproducibility, and low efficiency. In addition, the neurons obtained with these protocols are often immature and lack adequate functional activity both at the single-cell and network levels unless the neurons are cultured for several months. Partially due to these limitations, the functional properties of hiPSC-derived neuronal networks are still not well characterized. Here, we adapt a recently published protocol that describes production of human neurons from hiPSCs by forced expression of the transcription factor neurogenin-2 12. This protocol is rapid (yielding mature neurons within 3 weeks) and efficient, with nearly 100% conversion efficiency of transduced cells (>95% of DAPI-positive cells are MAP2 positive). Furthermore, the protocol yields a homogeneous population of excitatory neurons that would allow the investigation of celltype specific contributions to neurological disorders. We modified the original protocol by generating stably transduced hiPSC cells, giving us explicit control over the total number of neurons. These cells are then used to generate hiPSC-derived neuronal networks on micro-electrode arrays. In this way, the spontaneous electrophysiological activity of hiPSC-derived neuronal networks can be measured and characterized, while retaining interexperimental consistency in terms of cell density. The presented protocol is broadly applicable, especially for mechanistic and pharmacological studies on human neuronal networks.

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“…In the past decade, several protocols to convert hiPSCs into neurons have been developed [2][3][4][5][6][7][8] . However, these protocols are still limited in many ways.…”

Section: Introductionmentioning

confidence: 99%

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

Frega

Gestel

Linda

et al. 2017

JoVE

115

189

View full text Add to dashboard Cite

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“…A major advance from traditional differentiation methods is the circumvention of embryoid body (EB) formation for more efficient and direct induction of neural progenitor cells (NPCs) and expansion of neurospheres [99][100][101][102]. For example, Lie et al proffers high yield production of NPCs from feeder-free iPSC aggregates cultured in mTeSR™1(Stem Cell Technologies) [99].…”

Section: Neural Differentiation Of Ipscs: Quality and Quantitymentioning

confidence: 99%

“…A more protracted method by Shi et al induces iPSCs over 90 days to excitatory "cortical projection" neurons, with intermediate "cortical primary" stem/progenitor cells formed within 2 weeks, followed by "early-born" neurons produced between 2-3 weeks, and "last-born" neurons arising as late as day 90 [100]. The method is based on a much earlier protocol of SMAD signalling inhibition [93], and is purportedly highly efficient and less variable among different cell lines due to replacing noggin with SMAD inhibitor dorsomorphin [100].…”

Section: Neural Differentiation Of Ipscs: Quality and Quantitymentioning

confidence: 99%

“…In spite of the above mentioned challenges, improvements for iPSC differentiation to neural cells and tissues are being made through the development of better defined, optimised and efficient protocols[30, [99][100][101][102][103], bolstered by increased availability of superior stem cells attributable to improved somatic cell reprogramming, stem cell culture, banking and distribution [104][105][106][107]. A major advance from traditional differentiation methods is the circumvention of embryoid body (EB) formation for more efficient and direct induction of neural progenitor cells (NPCs) and expansion of neurospheres [99][100][101][102].…”

Section: Neural Differentiation Of Ipscs: Quality and Quantitymentioning

confidence: 99%

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The potential of induced pluripotent stem cells in models of neurological disorders: implications on future therapy

Crook¹,

Wallace

Tomaskovic-Crook

2015

Expert Review of Neurotherapeutics

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Tomaskovic-Crook, E. (2015). The potential of induced pluripotent stem cells in models of neurological disorders: Implications on future therapy. Expert Review of Neurotherapeutics: a key contribution to decision making in the treatment of neurologic and neuropsychiatric disorders, 15 (3), 295-304. Expert Review of NeurotherapeuticsThe potential of induced pluripotent stem cells in models of neurological disorders: Implications on future therapy AbstractThere is an urgent need for new and advanced approaches to modeling the pathological mechanisms of complex human neurological disorders. This is underscored by the decline in pharmaceutical research and development efficiency resulting in a relative decrease in new drug launches in the last several decades. Induced pluripotent stem cells represent a new tool to overcome many of the shortcomings of conventional methods, enabling live human neural cell modeling of complex conditions relating to aberrant neurodevelopment, such as schizophrenia, epilepsy and autism as well as age-associated neurodegeneration. This review considers the current status of induced pluripotent stem cell-based modeling of neurological disorders, canvassing proven and putative advantages, current constraints, and future prospects of nextgeneration culture systems for biomedical research and translation. There is an urgent need for new and advanced approaches to modelling the pathological

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“…Lou et al (2007) demonstrated that such stem cells contributed in the regeneration of damaged cells in the rat. Likewise, recovery of hair cells in vitro have been demonstrated in mice by application of fetal cochlea stem cells (Shi et al 2012;Savary et al 2007;Oshima et al 2007;White et al 2006;Doetzlhofer et al 2004). Also, embryonic and neural stem cells have been used to experimentally treat deafness in mice (Parker et al 2007; Corrales et al 2006).…”

Section: Introductionmentioning

confidence: 99%

Cochlear epithelial of dog fetuses: a new source of multipotent stem cells

et al. 2017

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Hearing loss caused by the damage of cochlea sensory cells or neurons is a common human disease, but also affects dogs and other animals. To test their progenitor nature as potential value for future therapies, we characterized cells derived from the cochlear epithelium in dog fetuses. In total, 8 fetuses of 35-40 days of gestation, derived from castration campaigns, were investigated. Cells were analysed by the MTT colorimetric assay and in regard to cell cycle, differentiation capacities, immunophenotypes and qPCR analysis. In culture, cells had a fibroblast-like morphology. Phenotypic immunocharacterization showed positive staining for mesenchymal stem cell and pluripotency markers and were negative for hematopoietic cell markers. Cells possessed differentiation capacity for the three main cell lineages: osteogenic, adipogenic and chondrogenic, altogether indicating their nature as mesenchymal stem cells. Thus, cells derived from fetal cochlear tissues indeed may provide valuable sources of progenitor cells for cell therapy of canine deafness and other diseases.

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Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks

Cited by 807 publications

References 24 publications

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays

The potential of induced pluripotent stem cells in models of neurological disorders: implications on future therapy

Cochlear epithelial of dog fetuses: a new source of multipotent stem cells

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