hN2™ Human Neural Discovery Kit

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Catalog: #hN27012.2.D | US $446 | International $650

The hN2™ Neural Discovery Kit is ideal for pilot experiments and assay optimization.  The kit contains the differentiated neural cells and optimized medium plus neural supplement for maintaining the cells in culture.  

hN2™ Differentiated Human Neuronal Cells are derived from the WA09 human embryonic stem cells by ArunA Biomedical using a proprietary process under defined feeder & serum free conditions. After differentiation, the cells exhibit a characteristic neuronal morphology and are highly post-mitotic. The cells are >90% β-III tubulin positive, >60% MAP2 positive, <5% Oct-4 positive and express high levels of glutamatergic and GABA-ergic phenotypes. These differentiated neural cells are  cryopreserved for ready, convenient use, therefore eliminating lengthy differentiation procedures by the end user.

hN2™ Differentiated Human Neuronal Cells can be cultured in an adherent 96-well format with ArunA’s proprietary culture medium and neural supplement and can be maintained in culture for >2 weeks during which time we have shown them to demonstrate extensive neurite outgrowth and synapse formation.

 

Time lapse image of hN2™ Differentiated Human Neuronal Cells,
for 24 hours in AB2™ Basal Neural Medium and ANS™ Neural Medium Supplement  based medium.
 

  

Cell Type:  neural cells
Part Number:  7012.2.D
Species:  human
Source:  derived from human embryonic cell line, H9 (WA09)
Morphology:  neuronal
Quantity:  ~1x106 cells per vial (viable post-thaw)
Shipping method:  dry ice
Storage:  liquid nitrogen
Growth Properties:  adherent, non-proliferative
Medium AB2™ Basal Neural Medium with ANS™ Neural Medium Supplement plus LIF (10 mg/mL)
Phenotyping:  Cells are >90% β III tubulin positive and >60% MAP2 positive.

DNA fingerprint:  Results are consistent with the presence of a single cell line with no evidence of culture cross contamination. The loci match the DNA fingerprint pattern for the H9 (NIH designation, WA09) hESC line as published in http://stemcells.nih.gov/research/nihresearch/scunit/.

Pathogen testing:   This lot was derived from the H9 hESC line that has been tested for Hepatitis B, Hepatitis C, HIV-1, HIV-2, HTLV-I/II, HSV1, HSV2, EBV, and CMV.  The H9 cell line has been tested and shown to be negative (performed by GIVF Laboratories).  Cells are tested for sterility and mycoplasma contamination.

 Kit Formats:

hN2™ Human Neural Discovery Kit (Cat# hN27012.2.D)

  • hN2™ Differentiated Human Neuronal Cells, 1 vial (~106 viable cells per vial)
  • AB2™ Basal Neural Medium, 1x 500-mL bottle (part 7011.3)
  • ANS™ Neural Medium Supplement, 1x 5-mL vial (part 7011.4) 

Technical Information:

Product Insert for hN2™ Human Neural Discovery Kit (PDF)
 

hN2™ Differentiated Human Neuronal Cells may be used in a wide variety of basic research applications such as cellular and molecular neurobiology, drug discovery, and neurotoxicity.

 Applications for the hN2™ Differentiated Human Neuronal Cells include studies of:

Neural stem cell references and reviews can be found here.

Product Versatility:

Human Neurotoxicity Screening

Toxicological HTS screening for neurobiological targets using hNP1™ Human Neural Progenitor Cells or hN2™ Differentiated Human Neuronal Cells has now become easier, faster, and standardized thanks to HemoGenix® Bioluminomics technology.

  • Excellent signal-to-noise ratio makes this an ideal choice for cell proliferation and cellular metabolism assays.
  • 96-or 384-well plate high throughput formats.
  • Sensitive, non-radioactive signal detection readout.

Fig. 1. Differences in cellular ATP levels between hNP1™ Human Neural Progenitor Cells & hN2™ Differentiated Human Neuronal Cells indicate differences in cell proliferation.

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Neurite Outgrowth

In conjunction with Molecular Devices' ImageeXpress HCI platform, hN2™ Differentiated Human Neuronal cells can be used to evaluate potential toxic effects of test compounds on nuerite outgrowth through visualizing cells stained with β-lll tubulin.

Fig. 1. Dose response curves for the effect of neurotoxic agents on neurite outgrowth. hN2™ cells were cultured in the presence of cytotoxic compounds for 72h, stained for β-lll tubulin, imaged with the ImageeXpress and analyzed for neurite outgrowth.

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Neural Progenitor Proliferation

Using hNP1™ Human Neural Progenitors Cells, Molecular Devices' ImageXpress platform can be used to quanitatively measure stimulators and inhibitors of neural progenitor proliferation through high content imaging (HCI). For data generated using this assay, please click here (PDF).

Fig. 1. Overlayed images from wells containing hNP1™ human neural progenitors and hN2™ differentiated human neuronal cells. Cells were stained with Hoeschst (nuclei; blue) and beta-III-tubulin (cytoskeleton; green). Images were acquired with ImageXpress Mircro system using a 20X objective.

Fig. 2. Image analysis results using Molecular Devices' MexaXpress Software (Cell Scoring Module) to measure the effects of growth factors on hNP1™ human neural progenitor proliferation (number of cells per field).

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Products are to be used for research purposes only, not for use in diagnostic or therapeutic applications. Products may not be sold, modified for sale or used for commercial purposes.

Frequently Asked Scientific Questions for hN2™ Differentiated Human Neuronal Cells

1. Are ArunA’s human cell lines derived from neural blastomas or cancerous cells?

The hN2™ Differentiated Human Neuronal Cells are derived from human embryonic stem cells that are karyotypically stable.

2. Do you have information about the transfection efficiency?

We have not tested transfection or transduction with the hN2™ Differentiated Human Neuronal Cells.

3. Are adult differentiated neurons electrophysiologically active?

Yes, our hN2™ Differentiated Human Neuronal Cells express a variety of ion channels and show some level of electrical activity.  A summary of ion channel gene expression and electrophysiological data for the hN2™ Differentiated Human Neuronal Cells is available upon request. Moreover, the hN2™ Differentiated Human Neuronal Cells form an adherent monolayer, making cell-based applications such as calcium imaging, electrophysiology and high content imaging much more straightforward than with neurospheres.

4. What are the phenotypes of your differentiated neural cells?

Our ready-to-use hN2™ Differentiated Human Neuronal Cells are derived using a proprietary differentiation process.  Validation standards include: expression of beta-III tubulin (>90% positive) and MAP2 (>60% positive); the cells are mostly post-mitotic. Although we do not routinely test for specific neuronal types, we have found that our process produces ~60% glutamatergic and ~40% GABA-ergic cells with <10% for other cell types (e.g, dopaminergic, serotonergic, cholinergic, astrocytic, etc.).

5. There is a lot of cell death/floating cells in my culture. Is this normal?

The hN2™
Differentiated Human Neuronal Cells are loosely adherent and it is not uncommon to have cells and cell debris floating in the culture media. You should allow for a 24-hour post-thaw recovery.  s long as the adherent cells are growing nicely in a monolayer on the plate, do not worry about the floating cells – they will be washed away during media changes. 

6.  How long can you keep the differentiated neural cells in culture?

hN2™ Differentiated Human Neuronal Cells may be maintained for up to two weeks in culture.  However, some researchers have reported maintaining hN2™ Differentiated Human Neuronal Cells for up to 4-5 weeks in culture.

7. Do the hN2™ Differentiated Human Neuronal Cells, when cultured, only express neuronal cell types?

In our hN2™ Differentiated Human Neuronal Cell cultures, we have observed a variety of neuronal phenotypes, especially after further exposure to additional growth factors used for selective enrichment.  Cell types include cholinergic, GABAergic and dopaminergic neurons.  Although the majority of the hN2™ Differentiated Human Neuronal Cell culture is comprised of neuronal cell types, astrocytes and oligodendrocytes have also been observed.  It's estimated that approximately 10% or less of the cells are glial.

If you need additional technical assistance, please contact us.

Products are to be used for research purposes only, not for use in diagnostic or therapeutic applications. It is prohibited to sell, modify for sale or use ArunA's products for commercial purposes without the express written consent of ArunA Biomedical.

ArunA has entered into a technology licensing agreement with the University of Georgia Research Foundation (UGARF) that enables the company to commercialize adherent monolayer neural cell technology developed at the University.  The license allows the company exclusive worldwide rights to develop and commercialize neural cells derived from human embryonic stem cells.

ArunA has also entered into a research and commercialization license with the Wisconsin Alumni Research Foundation (WARF) for nonexclusive rights to their embryonic stem cell patents in research applications.

 

Patent US 6,200,806

Patent US 7,531,354,B2 

Patent US 8,178,089

Left, Phase contrast image of hN2™ Differentiated Human Neuronal Cells in culture
Right, Confocal image of hN2™
Differentiated Human Neuronal Cellscells stained for the neuronal markers β III tubulin (green) and MAP2 (red) and for nuclei (blue).