Neural Precursor Cells

Gain a deeper understanding of the complexities of neuronal signaling and tease apart variations due to age, gender, and donor variability with neural precursor cells (NPCs) derived from human iPS cells.

  • Quick-Neuron™ Precursor - Human iPSC-derived Neural Precursor Cells (NPCs) give you access to the same cells with every order for more biologically relevant and reproducible studies
  • Quick-Neuron™ Precursor Kits provide you with a quick and easy way to turn your iPS cells into neural precursor cells (NPCs).
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iPSC-derived Neural Precursor Cells

Table of Contents

Advantages of iPSC-Derived Neural Precursor Cells

  • ~ 1 week differentiation
  • Complicated colony picking step NOT required
  • Highly pure population
  • No genetic footprint
  • Differentiation Kits and Differentiated Cells available
  • Multiple donors and disease types available

Characterization of iPSC-Derived Neural Precursor Cells

Differentiation kit workflow

NP-mRNA-S_workflow
NP-mRNA-S_phase-contrast

Representative phase contrast images of Quick-Neuron™ Precursor - mRNA Kit on days 1, 2, 3 and 7 days post-differentiation (scale bar = 100 μm). Cells are ready for passaging at Day 7.

iPSC-derived cells workflow

NP-mRNA-CW_workflow
iPSC-derived Neural Precursor Cell Phase Contrast images

Representative phase contrast images of Quick-Neuron™ Precursor – Human iPSC-derived Neural Precursor Cells on days 1-7 post-thaw (scale bar = 100 μm). Cells are ready for passaging at Day 7.

NP-mRNA-S_ICC

Immunofluorescent staining of Quick-Neuron™ Precursor - mRNA Kit shows expression of NES and VIM on day 7 post-differentiation (scale bar = 100 μm). Staining conditions: Anti-Nestin Antibody, clone 10C2 (Millipore Sigma, Catalog Number: MAB5326, 1:500 dilution) was used in combination with a secondary antibody (Invitrogen, Catalog Number: A11032 Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594, 1:500 dilution). Recombinant Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (abcam, Catalog Number: ab92547, 1:500 dilution) was used in combination with a secondary antibody (Invitrogen, Catalog Number: A32731 Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488, 1:500 dilution). Nuclei were counterstained with Hoechst 33342.

iPSC-derived Neural Precursor Cell ICC images

Immunofluorescent staining of Quick-Neuron™ Precursor – Human iPSC-derived NPCs show expression of nestin and vimentin on day 13 post-thaw (scale bar = 100 μm). Staining conditions:  Anti-Nestin Antibody, clone 10C2 (Millipore Sigma, Catalog Number: MAB5326, 1:500 dilution) was used  in combination with a secondary antibody (Invitrogen, Catalog Number: A11032 Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594, 1:500 dilution).  Recombinant Anti-Vimentin antibody [EPR3776] – Cytoskeleton Marker (abcam, Catalog Number: ab92547, 1:500 dilution) was used in combination with a secondary antibody (Invitrogen, Catalog Number: A32731 Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488, 1:500 dilution). Nuclei were counterstained with Hoechst 33342.

NP-mRNA_qPCR

Real-time quantitative PCR analysis of expression levels of neuronal precursor-associated genes NES, VIM, HES5, SOX1 and SOX2, as well as the stem cell pluripotency marker POU Class 5 Homeobox 1 (POU5F1/OCT4), were examined. Graph shows gene expression in Quick-Neuron™ Precursor Culture on day 6. The relative gene expression is normalized to phosphoglycerate kinase 1 (PGK1), and then calculated as a fold induction relative to undifferentiated hiPSCs as a control. Error bars show standard deviation.

iPSC-derived Neural Precursor Cell FACS

Flow cytometry analysis of NPC markers SOX2 and Nestin in Quick-Neuron™ Precursor cells at day 8 of differentiation. Cells were fixed in 4% PFA, permeabilized with BD Phosflow™ Perm Buffer III (cat# 558050) and then stained with either Alexa Fluor 488 Mouse-anti SOX2 (cat#561593) or Alexa Fluor 488 Mouse IgG1 κ Isotype Control (cat# 557721); shown in histogram A. Histogram B shows staining with either Alexa Fluor 647 Mouse-anti Nestin (cat# 560393) or Alexa Fluor 647 Mouse IgG1 κ Isotype Control (cat# 557732). Plots were derived from gated events on forward and side light scatter from an unstained NPC sample. Dot plot (C) demonstrates that >98% of the Quick-Neuron™ Precursor cells in culture are positive for SOX2 and Nestin. All antibodies were purchased from BD Biosciences.

Neural Precursor Cell Differentiation Kits

Our Quick-Neuron™ Precursor - mRNA Kits (small and large sizes available) allow researchers to quickly, easily, and efficiently differentiate their iPS or ES cell line of choice into neural precursor cells (NPCs). The kits utilize synthetic messenger RNA (syn-mRNA) to deliver our proprietary cocktail of transcription factors that induce differentiation without leaving a genetic footprint.

Quick-Neuron™ Precursor - mRNA Kits are available by request. Please inquire via email!

iPSC-Derived Neural Precursor Cells (NPCs)

Elixirgen Scientific’s Quick-Neuron™ Precursor - Human iPSC-derived Neural Precursor Cells are created using human iPS cells made available through a license from The California Institute for Regenerative Medicine (CIRM).

The CIRM iPS cell repository includes over 1,500 iPS cell lines, so please contact us if you would like neural precursor cells derived from a specific donor cell line. Every lot of iPSCs is differentiated into neural precursor cells using our proprietary transcription factor-based technology that is “footprint-free” and delivers functional, high-quality cells.

Quick-Neuron™ Precursor - Human iPSC-derived Neural Precursor Cells are available by request. Please inquire via email!

Frequently Asked Questions

Do I need a license to use Elixirgen’s iPSC-derived cells and/or differentiation reagent kits?

No. The use of differentiated cells or kits provided by Elixirgen Scientific does not require any additional license from other parties for any type of use, except for use in humans or for therapeutic or diagnostic use. Check out our license statement in Resource section for further detail.

What sizes of Quick-Neuron™ Excitatory - Human iPSC-derived Neurons are available?

Off the shelf, we offer small (>1 million viable cryopreserved cells) and large (5x >1 million viable cryopreserved cells) sizes. If you are looking for a larger quantity, please contact us for pricing information.

Should I use an mRNA- or SeV-based differentiation kit?

Both delivery methods provide robust “footprint-free,” non-integrating expression of the transcription factors, although each kit uses a slightly different workflow. Based on our data, the differentiation efficiency between an mRNA-based and a SeV-based kit is similar. For example, the percentages of TUBB3+ neurons differentiated using Quick-Neuron™ Cholinergic mRNA- and SeV-based kits were 93% and 88%, respectively.

We recommend the mRNA-based neuron differentiation kits for labs with experience culturing iPSCs/ESCs, as the workflow involves slightly more manipulation of fragile differentiating cells than the SeV-based kits. The SeV-based kits are better for labs with little experience culturing iPSCs/ESCs, although they must be used with BSL-2 safety measures and may require prior approval from your Institutional Biosafety Committee (IBC).

What sizes of Quick-Neuron™ differentiation kits are available?

Off the shelf, we offer small and large iPSC neuronal differentiation kits. The small size is best suited for single cell, imaging analyses, and pilot studies while the large size is best for applications such as high throughput screening and RNA sequencing.

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