iPSC Derived Human Neural Progenitors

Potent, Pure and Easy to Culture

We are pleased to announce the addition of Human Neural Progenitors to our Primary and Stem Cell offering.

Human Neural Progenitors at 95% Confluency

Cell potency, for us, includes the how well our cells can be differentiated into terminal types. For these progenitors, we have protocols for differentiating into neurons, astrocytes, and oligodendrocytes.

Neural Progenitors differentiated into Neurons and Stained with Tuj-1

We also have Neural Progenitors from Alcohol and Opioid-Addicted Donors.

Electrical Preconditioning of Stem Cells

Cool Science

The ability to manipulate hNPCs via a conductive scaffold creates a new approach to optimize stem cell-based therapy and determine which factors (such as VEGF-A) are essential for stroke recovery: Paul M. Georgea, Tonya M. Blissb, Thuy Huab, Alex Leed, Byeongtaek Oh, Alexa Levinson, Swapnil Mehta, Guohua Sun, Gary K. Steinberg. Electrical preconditioning of stem cells with a conductive polymer scaffold enhances stroke recovery. doi.org/10.1016/j.biomaterials.2017.07.020...anti βIII-tubulin (1:500, Neuromics, Edina, MN)...

Image: Neural progenitors were labeled with anti-rat Nestin polyclonal antibody (Cat#:GT15114) and stained with conjugated donkey anti-goat secondary anti-body (green). Differentiated neurons were labeled with neuron-specific mouse anti-β-III tubulin/ Tuj1-(Cat#MO15013 monoclonal antibody (red). Nuclei were stained with DAPI (blue).

We have excellent Stem Cell Differentiation Markers. Check them out.

Modeling HIV Latency

Generation of Infected Neurons
Researchers have developed a Neuronal Cell Line for the study of HSV-1 infection in humans. This line was developed by terminally differentiating human embryonic stem cells to neurons.

Our mouse monoclonal nestin antibody was used as a marker for the neural progenitor phase of this differentiation. Aldo Pourchet, Aram S. Modrek, Dimitris G. Placantonakis, Ian Mohr and Angus C. Wilson. Modeling HSV-1 Latency in Human Embryonic Stem Cell-Derived Neurons. Pathogens 2017, 6(2), 24; doi:10.3390/pathogens6020024.

Image:  In vitro derivation of human neural stem cells by differentiation of the Hes5::GFP human embryonic stem cell line. (A) Schematic showing the multistep neural induction protocol. TGFβi stands for TGF-β receptor I inhibitor (B) Bright field image of human embryonic stem cell (hESC) colonies cultured on mouse embryonic fibroblasts prior to reaching confluence. (C) Bright field image of rosette NSCs derived from dissociated hESC colonies cultured in neural induction media. (D) Phase contrast and indirect immunofluorescence images of NSC cultures grown on poly-l-ornithine/laminin-coated dishes in neural stem cell media and probed with an antibody against nestin, a neural stem cell marker. Nuclei were visualized with DAPI.

We will continue to post publications referencing use of our solutions.

Holistic Medicine for the Brain

Mind Expansion? 

There is a slowly growing body of support for certain hallucinogenic serotonin-like molecules supporting cognitive gains, antidepressant effects and changes in brain areas related to attention, self-referential thought, and internal mentation. These molecules are present in Virola Ayahuasca and used in traditional native South American medicine.

Here researchers found that in silico systems biology analyses support 5- MeO-DMT’s anti-inflammatory effects and reveal a modulation of proteins associated with the formation of dendritic spines, including proteins involved in cellular protrusion formation, microtubule dynamics and cytoskeletal reorganization. Proteins involved in long-term potentiation were modulated in a complex manner, with significant increases in the levels of NMDAR, CaMKII and CREB, but a reduction of PKA and PKC levels. These results offer possible mechanistic insights into the neuropsychological changes caused by the ingestion of substances rich in dimethyltryptamines: Vanja Dakic, Juliana Minardi Nascimento, Rafaela Costa Sartore, Renata de Moraes Maciel, Draulio B. de Araujo, Sidarta Ribeiro, Daniel Martins-de-Souza , Stevens Rehen. Short term changes in the proteome of human cerebral organoids induced by 5-methoxy-N,N-dimethyltryptamine. bioRxiv preprint first posted online Feb. 13, 2017; doi: http://dx.doi.org/10.1101/108159... anti-5-HT2A (RA24288, Neuromics)...

Images: Cerebral organoids express 5-MeO-DMT receptors and different cell type markers (A) Cerebral organoids presenting smooth texture and homogeneous coloring at 45 days of differentiation (scale bar 1000 µm). (B) Cerebral organoids are not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license. bioRxiv preprint first posted online Feb. 13, 2017; doi: http://dx.doi.org/10.1101/108159. The copyright holder for this preprint (which was 8 composed by several cell types, including mature neurons, as shown by MAP2 staining. (C) Cells expressing AMPAR1 are found in the organoid edge, while (D) cells expressing NMDAR1 and (E) GFAP are detected within the organoid. (F) Cells positive for 5-HT2A receptor, and (G) σ-1R, the primary molecular targets for 5-MeODMT, are also found in the organoid. Scale bars: A = 1000 µm; B = 50 µm; C, D, E, F, and G = 20 µm. (H) The expression of molecular targets for 5-MeO-DMT was also confirmed by RT-PCR.

Dimethyltryptamines should be studied further as potential therapies for depression and related disorders.

In Depth Analysis of Differentiating Neural Stem Cells

Foundation for Future Therapies

Here we share an example of this analysis. Results from worldwide research is now being made available via hPSCreg: Ilyas Singec, Andrew M. Crain, Junjie Hou, Brian T.D. Tobe, Maria Talantova, Alicia A. Winquist, Kutbuddin S. Doctor, Jennifer Choy, Xiayu Huang, Esther La Monaca, David M. Horn,5 Dieter A. Wolf, Stuart A. Lipton, Gustavo J. Gutierrez, Laurence M. Brill, and Evan Y. Snyder. Quantitative Analysis of Human Pluripotency and Neural Specification by In-Depth (Phospho)Proteomic Profiling. Stem Cell Reports (2016), http://dx.doi.org/10.1016/j.stemcr.2016.07.01.

Images: MDK Expression in Pluripotent and Differentiated Cells and Disruption of Neural Conversion by a Monoclonal Antibody against MDK (A) Representative immunocytochemical analysis of MDK expression by hESCs (WA09), skin fibroblasts (FB, line HS27), and fibroblasts (HS27) after reprogramming to hiPSCs. Scale bars represent 100 μm. (B) Western blot showing that MDK was not detected in HS27 fibroblasts but was expressed in WA09 hESCs and hiPSCs from HS27 fibroblasts, and was upregulated in differentiating cells (EBs and hNSCs from hESCs). Probing for GAPDH demonstrated similar loading of the lanes. (C) A monoclonal antibody against MDK did not impair pluripotent self-renewal of hESCs. Antibody concentrations are shown. (D) The percentage of pluripotent cells expressing OCT4 and NANOG did not differ significantly between anti-MDK and isotype control antibody treatment (3.0 μg/mL). Error bars represent SEM; n = 5. (E) qRT-PCR showed that a monoclonal antibody against MDK (1.5 μg/mL) inhibits neural gene expression during 6-day DAP treatment. Error bars represent mean + SEM, n = 3. ∗p less than  0.05. (F) Western blot analysis confirmed the inhibitory effect of anti-MDK on early neural marker expression (OTX1, PAX6), despite DAP treatment, in contrast to the isotype control. (G) Immunocytochemical evidence that neural conversion is inhibited by the anti-MDK antibody (1.5 μg/mL) during 6-day DAP treatment. Few cells induced PAX6 expression in the presence of anti-MDK antibody (right panel). Scale bar represents 200 μm. (H) Western blot analysis and time course of hESCs differentiated for 6 days with recombinant MDK alone (100 ng/mL) or a combination of the DAP cocktail and MDK (25 or 100 ng/mL) suggest that MDK specifically promotes neural commitment of hPSCs. SOX17, MIXL1, and Brachyury were not induced by MDK. In contrast, controls treated with fetal bovine serum (FBS) for 6 days, which stimulates multi-lineage differentiation of hPSCs, induced these non-neural markers.

This level of basic research yield knowledge that is important for the development of future cell based therapies for Neuro-related Diseases. We will continue to post updates here.

Human Astrocytes

Cortex Derived Astrocytes

Neuromics is pleased to be offering yet another option for culturing Human Astrocytes. They can be passaged up to 10X-Only 749 USD/500,000 Cells

They are deigned to be easy to culture and grow.

 Image: Human Brain Astrocytes cultured with our AlphaBioCoat.

Check out our large offering of Neurons, Astroglia, Progenitors, Brain Endothelial Cells/Pericytes and Blood Brain Barrier (BBB) Model. Should you have questions on these or any of our offerings, I can be reached directly at 612-801-1007 or pshuster@neuromics.com. Thank you, Pete Shuster-CEO and Owner.

Nestin Expression in Differentiating Neural Progenitors

Breathtaking Images

Our Nestin Antibody + a ​HES5::eGFP reporter was one of the markers used to visualize differentiating stem/progenitor cells (see: Nature Communications 6, Article number: 6500 doi:10.1038/ncomms7500).

Figures: (a) Neural differentiation scheme. Neural induction was performed by a dual SMAD inhibition protocol followed by long-term propagation with the factors indicated for 220 days. Naming conventions representing neuroepithelial (NE), early radial glial (E-RG), midradial glial (M-RG), late radial glial (L-RG) and long-term cultured progenitors (LNP) are indicated. Number of passages are indicated as P(n). (b) Bright field microscopy of progenitor cells during long-term differentiation shows dynamic morphological features. Scale bar: 50 μm (valid for all images in b). (c) Combined ​HES5::eGFP reporter expression and Nestin Immunostainings of stem/progenitor cells.

We have a large catalog of  Neuronal-Glial Markers. They are research proven and frequently published. Should have questions do not hesitate to call me directly 612-801-1007 or pshuster@neuromics.com. Pete Shuster, CEO and Owner.

Neuromics-ArunA Biomedical Partnership

We are Closer than Ever!

I spent the past several days meeting with Ms. Tracey Stice (COO), Dr. Steven Stice (President and CSO) and Ms. Joy Clark (Sales and Marketing Manager) at ArunA HQ in Athens, GA. The objective of the meeting was to finalize plans for our Strategic Selling Partnership. This partnership enables us to open up more cell based assay solutions to current and future Customers involved in neuro-diseases/disorders research and drug discovery.

The Aruna Team

...And the news gets better. By combining our expertise and experience, we are now well positioned to to:

1. Shape new solutions and services that would more tightly aligned with your unique requirements
2. Communicate how other labs within your organization are using our solutions and look for ways to leverage this.
3. Track buying volume so we can give favorable pricing based on purchase made in total by your organization vs specific groups.

We will be working even harder to make sure you that you are satisfied with the value we bring to you and your team. Wondering what we can do for you today? You can contact me directly 612-801-1007 or pshuster@neuromics.com. Thank you. Pete Shuster, CEO, Neuromics AND ArunA Biomedical Strategic Selling Partner.

Potents Tools for Neuroscience Based Toxicology Assays

Neuromics' Offers Best in Class Cell and Markers

I am always on the hunt for proof that are tools work in the many different applications required by Researchers Studying Neurotoxicology. Success is confirmed to us through Customer Data/Pubs and Testimonials.

I would like to feature here some examples:

• Neuron-Glial-Astrocyte-Progentitor Markers-Bin Xing, Adam D. Bachstetter, Linda J. Van Eldik. Inhibition of Neuronal p38α, but not p38β MAPK, Provides Neuroprotection Against Three Different Neurotoxic Insults. Journal of Molecular Neuroscience. July 2014...Cells were fixed with 3.7 % formaldehyde containing 0.1 % Triton X-100 in PBS for 10 min at room temperature. After washing three times with PBS, the coverslips were incubated with blocking buffer (PBS containing 6 % goat serum, 3 % bovine serum albumin (fraction V), 0.1 % Triton X-100) for 30 min at room temperature. Primary chicken anti-MAP2 antibody (1:1,000, Neuromics, Cat. no. CH22103) was diluted in blocking buffer and incubated with the cells at room tem- perature for 2 h. For detection of MAP2 staining, the cells were incubated with secondary biotin SP-conjugated goat antichicken antibody (1:1,000, Jackson ImmunoResearch) for 1 h, followed by streptavidin Alexa Fluor® fluorescent 488 (1:1,000, Invitrogen) incubation in blocking buffer at room temperature for 1 h. Wide field fluorescent photomicro- graphs were obtained using a Nikon Eclipse Ti microscope with an Axiocam MRc5 digital camera (Carl Zeiss)...

Figures: Neurons stained with Neuromics' MAP2 antibody to determine Neurite Damage.

• Primary Neurons and Astrocytes-Mark RichardsChee Wee PhoonGwendoline Tze Wei GohEng Khuan SengXu Ming GuoCherine Mei Fong TanWoon-Khiong ChanJoel Mun Kin Lee. A New Class of Pluripotent Stem Cell Cytotoxic Small Molecules. Research Article | published 19 Mar 2014 | PLOS ONE 10.1371/journal.pone.0085039 ...Primary hN2™ Human Neurons (Neuromics) were grown in hN2™ human neuron culture media (Neuromics)..
• Xiugong Gao, Hsiuling Lin, Radharaman Ray, Prabhati Ray. Toxicogenomic Studies of Human Neural Cells Following Exposure to Organophosphorus Chemical Warfare Nerve Agent VX. Neurochemical Research. February 2013. ...Human hN2 neurons were obtained from Neuromics...

We guarantee results. If you would like to learn more, please contact me directly at pshuster@neuromics.com or direct phone line: 612-801-1007. Thank you.

STEMEZ hNP1 Neural Progenitors and Ion Channels

In my conversation with neuro-drug discover researchers, I am frequently being asked about the potential of using our STEMEZ(TM) hNP1 Human Neural Progenitors Expansion Kits for studying ion channels. How effective are these cells as a source for studying neurodegenerative diseases and for drug screening assays? There is good news.

When differentiated, these  neural progenitors express subunits of glutamatergic,  GABAergic, nicotinic, purinergic and transient receptor potential receptors. In addition, sodium  and calcium channel subunits were also expressed. Functionally, virtually all the NP cells exhibited delayed rectifier potassium channel currents and some differentiated cells exhibited  tetrodotoxin sensitive, voltage-dependent sodium channel current under whole-cell voltage clamp and action potentials could be elicited by current injection under whole-cell current clamp.  These results indicate that removing basic fibroblast growth factor from the neural progenitor cell cultures leads to a post-mitotic state, and also results in the capability to produce excitable cells that can generate action potentials. This is the first data demonstrating capabilitiesof these cells for ionotrophic receptor assays and ultimately for electrically active human neural cell assays for drug discovery.

Images: Glutamate receptor expression in hNP cells and differentiated hNP cells The expression of ionotropic glutamate receptors might also be an indicator of neuronal maturation. These receptors are composed of three distinct families: NMDA, kainate and AMPA receptors. The hNP cells and differentiated hNP cells cultured in the absence of bFGF for 2 weeks were analyzed for mRNA expression of subunits of each glutamate receptor subtype relative to hESCs. Significant increases (p<0.05) in Grin2b were seen in hNP cells (20 fold) and differentiated hNP cells (25 fold) relative to hESCs (Figure 3A). Additionally, Grin1 and Grin2d were significantly increased (p<0.05) only in differentiated hNP cells relative to hESCs, but not in undifferentiated hNP cells (Figure 3A). Of the kainate receptors, Grik4 and Grik5 were significantly (p<0.05) increased only in undifferentiated hNP cells relative to hESCs (Figure 3B); whereas, Grik2 was significantly (p<0.05) increased only in hNP cells where bFGF had been removed (Figure 3B). AMPA receptor subunits were also examined. Gria1 and Gria4 were up regulated in hNP cells relative to hESCs (Figure 3C). Two week differentiated hNP cells showed significant (p<0.05) up regulation of Gria2 and Gira4 relative to hESCs (Figure 3C). To determine if functional glutamate channels exist in differentiated hNP cells, calcium influx in response to AMPA, kainic acid or NMDA application was measured on hNP cells, 14 days after the removal of bFGF. Figure 3G indicates that NMDA could not depolarize differentiated or undifferentiated hNP cells enough to cause significant calcium influx above background. In contrast, AMPA and kainic acid can cause calcium influx which can be potentiated by AMPA receptor specific modulator, cyclothiazide (50 μM, Figure 3G).Calcium influx was detected in the presence of cyclothiazide in calcium activity as measured (Figure 3H).

Images: Sodium channel activity in differentiated hNP cells was measured using whole cell voltage clamp. 81 total hNP cells cultured in the absence of bFGF from 4 to 27 days were analyzed. Of these, 34 exhibited no fast inward currents in response to a step depolarization indicating the 348 absence of functional voltage gated sodium channels (Figure 4G). The remaining cells yielded between 0.04 - 1.5 nA of inward current in response to the step depolarization (Figures 4B and 4G). These currents inactivated rapidly in all cases (Figures 4B and 4C) and could be abolished with the addition of 1 μM TTX (n = 3 cells; Figure 4C). Voltage-dependent steady state inactivation (n = 11 cells; Figure 4D) and recovery from fast inactivation (n = 5 cells; Figure 4E) were also observed on several positive cells. A subset of these cells was subjected to current clamp and action potentials were elicited by current injection (n = 8 cells, Figure 4F). In support of this, increasing concentrations of a sodium channel activator veratridine in a FLIPR assay on differentiated hNP cells show an increasing calcium response (Figure 4H). This probably resulted from voltage-gated sodium channel depolarization of cells that subsequently allowed calcium influx through calcium channels. These data indicate that differentiation of hNP cells by removal of bFGF can lead to a neuronal cell that can generate action potentials and depolarize the cell. The 58% hit rate for voltage-gated sodium channel function (Figure 4G), does not reflect the true proportion of sodium channel positive cells in our differentiated hNP cells, but rather our ability to morphologically distinguish these cells from negative cells by eye. An example of the morphology of a sodium channel positive cell is shown in Figure 4A. The positive cells were phase bright with a few long processes.

H9-WA09 Derived Human Neural Progenitors and Neurons Pricing

Neuromics offers stable, potent and well characterized STEMEZ (TM) Human Neural Progenitor & Neuron Discovery Kits. These are derived from NIH registered human ES cell line H9 (WA09).

Details on the capabilities of these cells are detailed in a variety of publications. They work! As a result, I am trying to make it easier to justify using them when and where they are needed. In addition to having frequent updates on new references, methods and data, I want to offer the best pricing possible.

In researching pricing of cells derived from the same parental line and having similar characteristics, I noticed prices ranging from 995 to 2800 USD. Our pricing starts at 695 USD and we offer deeper discounting if required. I do not want price to be a barrier. Here's some sample data on these cells.

Images: Neural phenotypes derived from hN2 cell lines. (A) Phase contrast image of differentiated culture. (B) Network including post-mitotic motoneurons (HB9). (C) Cholinergic neuron. (D) Tuj-1 positive cells that are DAT-positive (dopamine transporter; closed arrow) and DAT-negative (open arrow). (E) Gabaergic neurons, inset illustrates GABA in axon, but not the dendrites (arrow).

G-protein Coupled Receptor Expression Patterns Are Altered as Human Embryonic Stem Details-From Poster Presented at Neuroscience 2008 by Dr. Steve Stice et al.
hN2 Cells-Electro Phys Data.

Related Products:

• All Stem Cell Research Reagents
• Primary Neurons and Astrocytes

I wish exciting and rewarding discoveries.