High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1

Qu, Qiuhao; Liu, Dong; Louis, Kathleen; Li, Xiangzhen; Yang, Hong; Sun, Qinyu; Crandall, Shane R.; Tsang, Stephanie Jean; Zhou, Jiaxi; Cox, Charles L.; Cheng, Jianjun; Wang, Fei

doi:10.1038/ncomms4449

Cited by 120 publications

(122 citation statements)

References 48 publications

(76 reference statements)

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“…Genes assessed include the motor neuronspecific transcription factor, homeobox gene (HB9) [55] and Islet 1 (ISL1; n = 4-6 independent cultures for hESC-derived cells, and n = 3-4 independent cultures for iPSC-derived cells). Both HB9 and ISL1 are essential for the formation of mature motor neurons [55,56]. After 21 days of differentiation, 46% of hESC-derived cells and 11% of iPSC-derived cells costained positive for microtubule-associated protein 2 (MAP2) and ISL1 and 51% of hESC-derived cells and 16% of iPSCderived cells costained positive for MAP2 and HB9.…”

Section: Fig 1 Generation Of Motor Neurons (Mn) From Human Neural Smentioning

confidence: 99%

Differentiation of Human Neural Stem Cells into Motor Neurons Stimulates Mitochondrial Biogenesis and Decreases Glycolytic Flux

O’Brien

Keeney

Bennett

2015

Stem Cells and Development

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Differentiation of human pluripotent stem cells (hPSCs) in vitro offers a way to study cell types that are not accessible in living patients. Previous research suggests that hPSCs generate ATP through anaerobic glycolysis, in contrast to mitochondrial oxidative phosphorylation (OXPHOS) in somatic cells; however, specialized cell types have not been assessed. To test if mitobiogenesis is increased during motor neuron differentiation, we differentiated human embryonic stem cell (hESC)-and induced pluripotent stem cell-derived human neural stem cells (hNSCs) into motor neurons. After 21 days of motor neuron differentiation, cells increased mRNA and protein levels of genes expressed by postmitotic spinal motor neurons. Electrophysiological analysis revealed voltage-gated currents characteristic of excitable cells and action potential formation. Quantitative PCR revealed an increase in peroxisome proliferator-activated receptor gamma coactivator 1-a (PGC-1a), an upstream regulator of transcription factors involved in mitobiogenesis, and several of its downstream targets in hESC-derived cultures. This correlated with an increase in protein expression of respiratory subunits, but no increase in protein reflecting mitochondrial mass in either cell type. Respiration analysis revealed a decrease in glycolytic flux in both cell types on day 21 (D21), suggesting a switch from glycolysis to OXPHOS. Collectively, our findings suggest that mitochondrial biogenesis, but not mitochondrial mass, is increased during differentiation of hNSCs into motor neurons. These findings help us to understand human motor neuron mitobiogenesis, a process impaired in amyotrophic lateral sclerosis, a neurodegenerative disease characterized by death of motor neurons in the brain and spinal cord.

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Section: Fig 1 Generation Of Motor Neurons (Mn) From Human Neural Smentioning

confidence: 99%

Differentiation of Human Neural Stem Cells into Motor Neurons Stimulates Mitochondrial Biogenesis and Decreases Glycolytic Flux

O’Brien

Keeney

Bennett

2015

Stem Cells and Development

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“…Using vitronectin and xeno-free differentiation culture medium, the hiPSCs were able to differentiate into motor neurons under xeno-free conditions, and expressed markers of motor neurons. This xeno-free culture system greatly reduces the risk of immune rejection after cell transplantation and provides an invaluable tool for drug discovery and regenerative medicine by generation of patient-specific and clinical-grade iPSCs [20,28].…”

Section: Discussionmentioning

confidence: 99%

“…For hiPSC applications, directed differentiation to specific lineages with high efficiency and purity is a critical step [20]. Stem cell-derived motor neurons are being increasingly used to model diseases such as spinal muscular atrophy and amyotrophic lateral sclerosis in vitro.…”

Section: Introductionmentioning

confidence: 99%

Derivation, Expansion, and Motor Neuron Differentiation of Human-Induced Pluripotent Stem Cells with Non-Integrating Episomal Vectors and a Defined Xenogeneic-free Culture System

Xiong

et al. 2015

Mol Neurobiol

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Induced pluripotent stem cells (iPSCs) generated from patient-derived somatic cells provides the opportunity for model development in order to study patient-specific disease states with the potential for drug discovery. However, use of lentivirus and exposure of iPSCs to animal-derived products limit their therapeutic utility and affect lineage differentiation and subsequent downstream functionality of iPSC derivatives. Within the context of this study, we describe a simple and practical protocol enabling the efficient reprogramming of terminally differentiated adult fibroblasts into integration-free human iPSCs (hiPSCs) using a combination of episomal plasmids with small molecules (SMs). Using this approach, there was a 10-fold increase in reprogramming efficiency over single plasmid vector-based methods. We obtained approximately 100 iPSCs colonies from 1 × 10(5) human adult dermal fibroblasts (HADFs) and achieved approximately 0.1% reprogramming efficiencies. Concurrently, we developed a highly conducive culture system using xeno-free media and human vitronectin. The resulting hiPSCs were free of DNA integration and had completely lost episomal vectors, maintained long-term self-renewal, featured a normal karyotype, expressed pluripotent stem cell markers, and possessed the capability of differentiating into components of all three germ layers in vivo. Finally, we demonstrate that the integration-free hiPSCs could be differentiated into motor neurons under xeno-free culture conditions. This induction method will promote the derivation of patient-specific integration-free and xeno-free iPSCs and improve the strategy for motor neuron derivation. Our approach provides a useful tool for human disease models, drug screen, and clinical applications.

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“…Thus, human umbilical cord mesenchymal stem cells may serve as an alternative source of multipotent stem cells for the treatment of neurodegenerative disease. Producing motor neurons through differentiating of ESCs, ASCs, and human adipose-derived stem cells (hADSCs) showed successful results (Wichterle et al 2002;Qu et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Induction of human umbilical Wharton’s jelly-derived mesenchymal stem cells toward motor neuron-like cells

Bagher

Ebrahimi‐Barough

Azami

et al. 2015

In Vitro Cell.Dev.Biol.-Animal

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The most important property of stem cells from different sources is the capacity to differentiate into various cells and tissue types. However, problems including contamination, normal karyotype, and ethical issues cause many limitations in obtaining and using these cells from different sources. The cells in Wharton's jelly region of umbilical cord represent a pool source of primitive cells with properties of mesenchymal stem cells (MSCs). The aim of this study was to determine the potential of human Wharton's jelly-derived mesenchymal stem cells (WJMSCs) for differentiation to motor neuron cells. WJMSCs were induced to differentiate into motor neuron-like cells by using different signaling molecules and neurotrophic factors in vitro. Differentiated neurons were then characterized for expression of motor neuron markers including nestin, PAX6, NF-H, Islet 1, HB9, and choline acetyl transferase (ChAT) by quantitative reverse transcription PCR and immunocytochemistry. Our results showed that differentiated WJMSCs could significantly express motor neuron biomarkers in RNA and protein levels 15 d post induction. These results suggested that WJMSCs can differentiate to motor neuron-like cells and might provide a potential source in cell therapy for neurodegenerative disease.

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High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1

Cited by 120 publications

References 48 publications

Differentiation of Human Neural Stem Cells into Motor Neurons Stimulates Mitochondrial Biogenesis and Decreases Glycolytic Flux

Differentiation of Human Neural Stem Cells into Motor Neurons Stimulates Mitochondrial Biogenesis and Decreases Glycolytic Flux

Derivation, Expansion, and Motor Neuron Differentiation of Human-Induced Pluripotent Stem Cells with Non-Integrating Episomal Vectors and a Defined Xenogeneic-free Culture System

Induction of human umbilical Wharton’s jelly-derived mesenchymal stem cells toward motor neuron-like cells

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