Large-Scale Production of Mature Neurons from Human Pluripotent Stem Cells in a Three-Dimensional Suspension Culture System

Autor: Lee L. Rubin, Karen Weisinger, Giuliana G. Repetti, Quinn P. Peterson, Alessandra Rigamonti, Feodor D. Price, Chen Benkler, Chicheng Sun, Lance S. Davidow, Emil M. Hansson, Francesca Rapino, Danielle C. Reny
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Nervous system
Resource
Pluripotent Stem Cells
Cellular pathology
Cellular differentiation
Neurogenesis
Cell Culture Techniques
Smad Proteins
Biology
Biochemistry
03 medical and health sciences
Spheroids
Cellular
Genetics
medicine
Humans
Ciliary Neurotrophic Factor
Glial Cell Line-Derived Neurotrophic Factor
Induced pluripotent stem cell
lcsh:QH301-705.5
Neurons
Observer Variation
lcsh:R5-920
Brain-Derived Neurotrophic Factor
Tumor Suppressor Proteins
Reproducibility of Results
Depolarization
Cell Differentiation
Cell Biology
Cell biology
Repressor Proteins
030104 developmental biology
medicine.anatomical_structure
lcsh:Biology (General)
Cell culture
Nerve Net
T-Box Domain Proteins
lcsh:Medicine (General)
Developmental biology
Microtubule-Associated Proteins
Biomarkers
Developmental Biology
Zdroj: Stem Cell Reports, Vol 6, Iss 6, Pp 993-1008 (2016)
Stem Cell Reports
ISSN: 2213-6711
Popis: Summary Human pluripotent stem cells (hPSCs) offer a renewable source of cells that can be expanded indefinitely and differentiated into virtually any type of cell in the human body, including neurons. This opens up unprecedented possibilities to study neuronal cell and developmental biology and cellular pathology of the nervous system, provides a platform for the screening of chemical libraries that affect these processes, and offers a potential source of transplantable cells for regenerative approaches to neurological disease. However, defining protocols that permit a large number and high yield of neurons has proved difficult. We present differentiation protocols for the generation of distinct subtypes of neurons in a highly reproducible manner, with minimal experiment-to-experiment variation. These neurons form synapses with neighboring cells, exhibit spontaneous electrical activity, and respond appropriately to depolarization. hPSC-derived neurons exhibit a high degree of maturation and survive in culture for up to 4–5 months, even without astrocyte feeder layers.
Highlights • Efficient, large-scale differentiation of hPSC to distinct neuronal subtypes • Differentiated neurons can be maintained in spinner flasks for extended time periods • Derived neurons are functionally mature and form an extensive synaptic network • Attractive platform for in vitro modeling of different forms of neurological disease
Ruben and colleagues present new protocols for large-scale production of cortical and motor neurons from human pluripotent stem cells. The obtained neurons are functional, survive an extended period of time in culture without the use of feeder layers, and constitute a promising platform for in vitro modeling of neurological diseases.
Databáze: OpenAIRE
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