hNP1 Neural Progenitors and Neuronal Regeneration after Injury

The Potential of Axonal PPARγ

In this study, researchers used our hNP1^TM Neural Progenitors to demonstrate that axonal PPARγ is involved in neuronal injury responses required for axonal regeneration: Juan Pablo Lezana, Shachar Y. Dagan, Ari Robinson, Ron S. Goldstein, Mike Fainzilber, Francisca C. Bronfman, and Miguel Bronfman. Axonal pparγ promotes neuronal regeneration after injury. DOI: 10.1002/dneu.22353... Culture and regeneration analysis of human axons. NP1 human neural precursors were purchased from Neuromics (Minnesota, USA) and passaged up to 8 times before generation of neurons. Cells were used up until passage 10 after defrosting the purchased neural...

It is important to note that the researchers built up a large stock by passaging the progenitors prior to differentiation.

Check out our hNP1 publications to learn of more unique applications!

Save 100 USD on our hN2™ Primary Neurons

Potent, Pure and Easy to Culture

Assay data continues to roll in. I am pleased by the high lot to lot consistencies of our hN2 Primary Human Neurons resulting in data reproducibility. Here're an examples of outgrowth/tox assays: In conjunction with Molecular Devices' ImageeXpress HCI platform, hN2 differentiated neuronal cells can be used to evaluate potential toxic effects of test compounds on nuerite outgrowth through visualizing cells stained with β-lll tubulin.

Figure: 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.

I am always available for product related issues and questions. I cam be reached at direct phone: 612-801-1007 and pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics.

Neurite Outgrowth Assays-96 Well Plates

Fast, Effective and Data Rich!

Our Customers' are reporting great results using our hN2 Primary Human Neurons. We are seeing a natural evolution for high content to high throughput screening. This is because our human neuron culturing kits are proving pure and potent (sample pubs).

Here's some customer data using hN2™ Human Neurons High Throughput Screening (HTS) Kit:

Image: Neurite Outgrowth Assay in 96 Well Plate using hN2 Neurons-Blue – Nuclear staining; Green – Neurite and cell body staining.

Want to learn more? Contact me directly at pshuster@neuromics.com or 612-801-1007. Thank you. Pete Shuster, CEO and Owner, Neuromics.

Good Axon; Bad Axon: Regeneration in the CNS and Age

"The incapacity of the central nervous pathways to regenerate is a dogma accepted by science..." - Ramon y Cajal


Harvard University has opened access to Michio Wendell Painter's Dissertation: Regeneration in the aging peripheral nervous system. Will Spinal Cord Injury (SCI) and Neurodegenerative Diseases of the PNS become treatable with regenerative therapies? This work provides important insights.

Age plays a central role in regenerative capacities.

Neurite Outgrowth Assays

Cells, Media and Markers

I have previously posted use of our Neurons in Live Content Assays for the study of Neurite Outgrowth: Neurons-Live Content Assays. These assays are critical for the study of repair and regeneration.

A recent publication featured several of our Neuron Markers: Serena Quarta, Bastian E. Baeumer, Nadja Scherbakov1, Manfred Andratsch, Stefan Rose-John, Georg Dechant3, Christine E. Bandtlow, and Michaela Kress: Peripheral Nerve Regeneration and NGF-Dependent Neurite Outgrowth of Adult Sensory Neurons Converge on STAT3 Phosphorylation Downstream of Neuropoietic Cytokine Receptor gp130. The Journal of Neuroscience, 24 September 2014, 34(39): 13222-13233; doi: 10.1523/JNEUROSCI.1209-13.2014.
Live labeling of neuron cultures: After 20 or 48 h, neurons were live-labeled with α-gp130 antibody diluted in cold TNB medium for 30 min on ice. After washing, neurons were incubated with the secondary antibody diluted in cold TNB medium for 30 min and washed with PBS. Cells were fixed either with 4% PFA for 20 min at room temperature (RT) or with methanol at −20°C for 2 min. After permeabilization with 0.01% TX-100 (Pierce) unspecific binding was blocked for 30 min with 10% normal goat serum (Sigma-Aldrich) in PBS. Cells were incubated with the first antibody for 1 h, washed three times for 10 min with PBS and incubated with the appropriate secondary antibody for 30 min, counterstained with 4′, 6-diamidino-2-phenylindole (1:10,000; Sigma-Aldrich) and embedded in Mowiol (Calbiochem). As primary antibodies, α-gp130 (1:50; Neuromics), α-β-III-tubulin clone TuJ-1 (1:1000; R&D Systems), and α-neurofilament-H (α-NF-H; 1:200; Neuromics) were used. Secondary antibodies used were α-goat AlexaFluor 594 (1:1000; Invitrogen), chicken α-mouse AlexaFluor 594 or donkey α-mouse AlexaFluor 488 (1:1000; Invitrogen), and goat α-chicken AlexaFluor 568 (1:10,000; Invitrogen) for fluorescence microcopy.

Images: Reduced density of TuJ-1+ nerve endings in the epidermis in SNS-gp130−/− mice after lesion. A, Representative cross sections of hindpaw glabrous skin of naive and 12 dpl gp130fl/fl and SNS-gp130−/− mice stained with the pan neuronal marker TuJ-1. The dotted line indicates the border between dermis and epidermis. Scale bar, 40 μm. B, Quantification of the total number of TuJ-1+ fibers (NE) per 1000 μm2 of epidermal area shows a significant decrease in density in SNS-gp130−/− mice after lesion compared with control animals (*p < 0.05; n = 4 for each group). Data are presented as mean ± SEM and analyzed by Mann–Whitney U test. C, 3D reconstruction of the deeper layer of the dermis shows fewer nerve bundles in SNS-gp130−/− dermis compared with controls. D, No NF-H+ proprioceptive fibers were detectable in the epidermis of gp130fl/fl animals at 12 dpl. Scale bar, 40 μm.

If you want to learn more about our Neuron-Glial-Astrocyte based assay solutions do not hesitate to contact me (612-801-1007) or pshuster@neuromics.com. Pete Shuster, Owner and CEO, Neuromics.

Neurons in 3-D

In Vivo Like Neurite Outgrowth Cultures

Culturing in 3-D requires potent Primary Neurons. Here researchers use our e18 Rat Cortical Neurons in developing their 3-D assays: Chandrasekhar R. Kothapallia, Peyman Honarmandib. Theoretical and experimental quantification of the role of diffusive chemogradients on neuritogenesis within 3D collagen scaffolds. Acta Biomaterialia. Available online 14 May 2014. http://dx.doi.org/10.1016/j.actbio.2014.05.009

Abstract: A critical challenge to regenerating close mimics of native axonal pathways under chronic neurodegenerative disease or injury conditions is the inability to stimulate, sustain and steer neurite outgrowth over a long distance, till they reach their intended targets. Understanding neurite outgrowth necessitates quantitative determination of the role of molecular gradients on growth cone navigation under dynamic physiological conditions. High-fidelity biomimetic platforms are needed to computationally and experimentally acquire and analyze spatio-temporal molecular gradient evolution and the growth cone response across multiple conditions along this gradient pathway. In this study, we utilized a simple microfluidic platform in which diffusive gradients were generated within a 3D porous scaffold in a defined and reproducible manner, and its characteristics (spatio-temporal gradient, steepness, diffusion time, etc.) precisely quantified at every specific location within the scaffold. Using this platform, we show that the cortical neurite response within 3D collagen scaffolds, at both the cellular and molecular level, is extremely sensitive to subtle changes in localized concentration and gradient steepness of IGF-1 within that region. This platform could also be used to study other biological processes such as morphogenesis and cancer metastasis, where chemogradients are expected to significantly regulate the outcomes. Results from this study might be of tremendous use in designing biomaterial scaffolds for neural tissue engineering, axonal pathway regeneration under injury or disease, and in formulating targeted drug delivery strategies.

Image: Neurons in 3-D Assay

Neuromics' provides many Stem and Primary Cell Assay Solutions including tools for 3-D Cultures. We also offer services studying the effects of small molecules and compounds on Stem Cell expansion, differentiation and migration. To learn more contact me at 612-801-1007 or pshuster@neuromics.com.

Live Cell Imaging

My friends at Essen Bioscience are taking live cell imaging to new heights. I previously posted their neurite outgrowth solutions (hippcampal neurons).

In this posting, I would like share highlights from the December 2012 issue of "Genetic and Biotechnology News": Essen BioScience’s IncuCyte ZOOM™(live cell imaging in your incubator) and CellPlayer™ reagents enable you to overcome the limitations of static cell based assays. The solutions enable the acquisition, analysis, and quantification of images from living cells that remain unperturbed by the detection method, allowing for repeated measures of cell biology over long periods of time, from days to weeks. This is true Live Content Imaging.

Example: Real-Time Cell Counting in Mixed Cultures.
Cell-based models composed of more than one cell type in the same culture are increasingly recognized as more biologically relevant than monocultures. For example, a recent cancer study illustrated that some stromal cell types confer resistance to tumor cells in coculture, proposing this as a possible mechanism
of tumor resistance in the clinic.

Integrated image processing algorithms provided an independent nuclear count of both cell types continuously in time. The combined attributes of this approach can be used to better elucidate the mechanism and timing of drug responses on cell proliferation in biologically relevant, mixed culture systems.

Capabilities extend to a wide range of other phenotypic assays, including cell death, angiogenesis, neurite dynamics, and cell migration and invasion.

These solutions are have the ability to save time and drive costs out of the drug discovery process. I will keep you posted.

Neuromics' Cortical Neurons & Kinetic NeuroTrack Assays

Providing tools that insure excellent Cell Based Assays is a cornerstone of our business strategy. Lauren McGillicuddy and her team at Essen Bioscience have been using our E18 Primary Rat Cortical Neurons to develop NeuroTrak^TM assays enabling kinetic quantification of neurite dynamics (initiation, branching, extension, retraction). NeuroTrack is one of several CellPlayer^TM assays that can be run in IncuCyte Zoom^TM.

The proof is in the results and these show the both the potency of the cells and the powerful capablities of the IncuCyte ZOOM hardware and software:

Images:  Neurite outgrowth of rat E18 cortical neurons in a 96-well microplate at 24, 48 and 120 hours. top: 20x HD phase image of primary neurons  bottom: Image segmentation of neurites (light blue) and cell body cluster (raspberry).

Images: Neurite outgrowth of rat E18 cortical neurons in a 96-well microplate. Left: 20x HD phase image of primary neurons, 96 hours post plating ; Middle: Image segmentation of neurites (yellow) and cell body cluster (raspberry); Right: Concentration and time dependent inhibition of neurite outgrowth with the protein kinase C inhibitor, Ro-31-8220 (mean ± SD; n=6 per condition).

Check out this cool  assay animation!

I will continue to keep you posted on new assays and related methods using our primary neuron and astroglial cells.

Enhancing Neurite Outgrowth

We are pleased to announce the addition of new proteins to our Axon Growth and Guidance category. These are designed to enhance neurite out growth.

Name	Catalog #	Type	Species	Size	Price
NGF-b, NSO Derived	PR15083	Protein	H; M; R	100 ug	$285
NGF-b, NSO Derived, CF	PR15084CF	Protein	H; M; R	100 ug	$285
S100A13, CF	PR15085CF-50	Protein	H; R	50 ug	$315
Slit1	PR15075-50	Protein	Ch; H	50 ug	$315
Slit2, CHO	PR15085-50	Protein	Ch; H	50 ug	$315

Image: Cultured chick dorsal root ganglion neurons were grown in the presence of recombinant human NGF-b with (A) or without (B) recombinant mouse Slit2. The presence of the Slit2 protein signifi cantly enhanced neurite outgrowth.