STEMdiff™ Neural Progenitor Medium

Medium for maintenance and expansion of neural progenitor cells derived from human ES and iPS cells

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STEMdiff™ Neural Progenitor Medium

Medium for maintenance and expansion of neural progenitor cells derived from human ES and iPS cells

1 Kit
Catalog #05833
343 USD

Overview

STEMdiff™ Neural Progenitor Medium is a defined and serum-free medium for the expansion of neural progenitor cells (NPCs) derived from human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells using STEMdiff™ Neural Induction Medium (Catalog #05835). NPCs cultured in this medium can be expanded 3-5 fold per passage, and cultured for at least 10 passages, with minimal spontaneous neuronal differentiation.
Advantages:
• Defined and serum-free
• Supports expansion of NPCs generated using STEMdiff™ Neural Induction Medium
• Optimized for efficient expansion of NPCs over multiple passages
• Preserves NPC multipotency while minimizing spontaneous neuronal differentiation
• Convenient, user-friendly format and protocol
Components:
  • STEMdiff™ Neural Progenitor Basal Medium, 500 mL
  • STEMdiff™ Neural Progenitor Supplement A (50X), 10 mL
  • STEMdiff™ Neural Progenitor Supplement B (1000X), 500 µL
Subtype:
Specialized Media
Cell Type:
Neural Cells, PSC-Derived; Neural Stem and Progenitor Cells; Pluripotent Stem Cells
Species:
Human
Application:
Cell Culture; Expansion
Brand:
STEMdiff
Area of Interest:
Disease Modeling; Drug Discovery and Toxicity Testing; Neuroscience; Stem Cell Biology
Formulation:
Serum-Free

Scientific Resources

Educational Materials

(17)
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Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Morphology and Marker Expression of Neural Progenitor Cells Cultured in STEMdiff™ Neural Progenitor Medium

Figure 1. Morphology and Marker Expression of Neural Progenitor Cells Cultured in STEMdiff™ Neural Progenitor Medium

(A) Typical NPC morphology is observed in cultures (shown at day 6 of passage 1). (B-D) NPCs maintained in STEMdiff™ Neural Progenitor Medium express the CNS-type NPC markers PAX6 (B, D, red), SOX1 (C, red) and NESTIN (C, green), but not the neural crest marker SOX10 (D, green, single channel shown in inset). B-D were taken at the same magnification.

Expansion of Neural Progenitor Cells in STEMdiff™ Neural Progenitor Medium

Figure 2. Expansion of Neural Progenitor Cells in STEMdiff™ Neural Progenitor Medium

NPCs cultured in STEMdiff™ Neural Progenitor Medium can be expanded to generate a large number of cells. Three- to five-fold expansion can be achieved upon each passage. NPCs were derived using STEMdiff™ Neural Induction Medium and passaged once a week on average. n = 6.

Neural Progenitor Cells Cultured in STEMdiff™ Neural Progenitor Medium Show Minimal Spontaneous Neuronal Differentiation

Figure 3. Neural Progenitor Cells Cultured in STEMdiff™ Neural Progenitor Medium Show Minimal Spontaneous Neuronal Differentiation

Passages 1 (A) and 3 (B) of a representative NPC culture maintained in STEMdiff™ Neural Progenitor Medium. Cells were immunolabeled with SOX1 (red) to identify NPCs, and class III β-tubulin (green) to identify neurons. Spontaneous neuronal differentiation is low in NPC cultures maintained in STEMdiff™ Neural Progenitor Medium. A and B were taken at the same magnification.

Neural Progenitor Cells Maintained in STEMdiff™ Neural Progenitor Medium can Differentiate into Neurons and Astrocytes

Figure 4. Neural Progenitor Cells Maintained in STEMdiff™ Neural Progenitor Medium can Differentiate into Neurons and Astrocytes

When directed according to published protocols, NPCs can differentiate into neurons (A, class III β-tubulin shown in red) and astrocytes (B, GFAP shown in red). Nuclei are counterstained with DAPI (blue).

Publications

(12)
Stem cell reports 2018 JUL

Disruption of GRIN2B Impairs Differentiation in Human Neurons.

S. Bell et al.

Abstract

Heterozygous loss-of-function mutations in GRIN2B, a subunit of the NMDA receptor, cause intellectual disability and language impairment. We developed clonal models of GRIN2B deletion and loss-of-function mutations in a region coding for the glutamate binding domain in human cells and generated neurons from a patient harboring a missense mutation in the same domain. Transcriptome analysis revealed extensive increases in genes associated with cell proliferation and decreases in genes associated with neuron differentiation, a result supported by extensive protein analyses. Using electrophysiology and calcium imaging, we demonstrate that NMDA receptors are present on neural progenitor cells and that human mutations in GRIN2B can impair calcium influx and membrane depolarization even in a presumed undifferentiated cell state, highlighting an important role for non-synaptic NMDA receptors. It may be this function, in part, which underlies the neurological disease observed in patients with GRIN2B mutations.
Neuroscience letters 2017 JAN

Generation of disease-specific autopsy-confirmed iPSCs lines from postmortem isolated Peripheral Blood Mononuclear Cells

Belle K et al.

Abstract

Understanding the molecular mechanisms that underlie neurodegenerative disorders has been hampered by a lack of readily available model systems that replicate the complexity of the human disease. Recent advances in stem cell technology have facilitated the derivation of patient-specific stem cells from a variety of differentiated cell types. These induced pluripotent stem cells (iPSCs) are attractive disease models since they can be grown and differentiated to produce large numbers of disease-relevant cell types. However, most iPSC lines are derived in advance of, and without the benefit of, neuropathological confirmation of the donor - the gold standard for many disease classifications and measurement of disease severity. While others have reported the generation of autopsy-confirmed iPSC lines from patient explants, these methods require outgrowth of cadaver tissue, which require additional time and is often only successul 50% of the time. Here we report the rapid generation of autopsy-confirmed iPSC lines from peripheral blood mononuclear cells (PBMCs) drawn postmortem. Since this approach doesn't require the propagation of previously frozen cadaver tissue, iPSC can be rapidly and efficiently produced from patients with autopsy-confirmed pathology. These matched iPSC-derived patient-specific neurons and postmortem brain tissue will support studies of specific mechanisms that drive the pathogenesis of neurodegenerative diseases.
Cell stem cell 2017 JAN

Recent Zika Virus Isolates Induce Premature Differentiation of Neural Progenitors in Human Brain Organoids.

E. Gabriel et al.

Abstract

The recent Zika virus (ZIKV) epidemic is associated with microcephaly in newborns. Although the connection between ZIKV and neurodevelopmental defects is widely recognized, the underlying mechanisms are poorly understood. Here we show that two recently isolated strains of ZIKV, an American strain from an infected fetal brain (FB-GWUH-2016) and a closely-related Asian strain (H/PF/2013), productively infect human iPSC-derived brain organoids. Both of these strains readily target to and replicate in proliferating ventricular zone (VZ) apical progenitors. The main phenotypic effect was premature differentiation of neural progenitors associated with centrosome perturbation, even during early stages of infection, leading to progenitor depletion, disruption of the VZ, impaired neurogenesis, and cortical thinning. The infection pattern and cellular outcome differ from those seen with the extensively passaged ZIKV strain MR766. The structural changes we see after infection with these more recently isolated viral strains closely resemble those seen in ZIKV-associated microcephaly.
Nucleic acids research 2017 FEB

A genome-integrated massively parallel reporter assay reveals DNA sequence determinants of cis-regulatory activity in neural cells.

Maricque BB et al.

Abstract

Recent large-scale genomics efforts to characterize the cis-regulatory sequences that orchestrate genome-wide expression patterns have produced impressive catalogues of putative regulatory elements. Most of these sequences have not been functionally tested, and our limited understanding of the non-coding genome prevents us from predicting which sequences are bona fide cis-regulatory elements. Recently, massively parallel reporter assays (MPRAs) have been deployed to measure the activity of putative cis-regulatory sequences in several biological contexts, each with specific advantages and distinct limitations. We developed LV-MPRA, a novel lentiviral-based, massively parallel reporter gene assay, to study the function of genome-integrated regulatory elements in any mammalian cell type; thus, making it possible to apply MPRAs in more biologically relevant contexts. We measured the activity of 2,600 sequences in U87 glioblastoma cells and human neural progenitor cells (hNPCs) and explored how regulatory activity is encoded in DNA sequence. We demonstrate that LV-MPRA can be applied to estimate the effects of local DNA sequence and regional chromatin on regulatory activity. Our data reveal that primary DNA sequence features, such as GC content and dinucleotide composition, accurately distinguish sequences with high activity from sequences with low activity in a full chromosomal context, and may also function in combination with different transcription factor binding sites to determine cell type specificity. We conclude that LV-MPRA will be an important tool for identifying cis-regulatory elements and stimulating new understanding about how the non-coding genome encodes information.
Biomaterials 2016 MAY

Simple and versatile synthetic polydopamine-based surface supports reprogramming of human somatic cells and long-term self-renewal of human pluripotent stem cells under defined conditions

Zhou P et al.

Abstract

Human pluripotent stem cells (hPSCs) possess great value in the aspect of cellular therapies due to its self-renewal and potential to differentiate into all somatic cell types. A few defined synthetic surfaces such as polymers and adhesive biological materials conjugated substrata were established for the self-renewal of hPSCs. However, none of them was effective in the generation of human induced pluripotent stem cells (hiPSCs) and long-term maintenance of multiple hPSCs, and most of them required complicated manufacturing processes. Polydopamine has good biocompatibility, is able to form a stable film on nearly all solid substrates surface, and can immobilize adhesive biomolecules. In this manuscript, a polydopamine-mediated surface was developed, which not only supported the reprogramming of human somatic cells into hiPSCs under defined conditions, but also sustained the growth of hiPSCs on diverse substrates. Moreover, the proliferation and pluripotency of hPSCs cultured on the surface were comparable to Matrigel for more than 20 passages. Besides, hPSCs were able to differentiate to cardiomyocytes and neural cells on the surface. This polydopamine-based synthetic surface represents a chemically-defined surface extensively applicable both for fundamental research and cell therapies of hPSCs.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.