hNP1 Neural Progenitors and HTS Tox Assays

Sensitivity of Neural Progenitor Cells to Chemical Induced Apoptosis

This article demonstrates the sensitivity of Neuromics' hNP1TM Neural Progenitors to chemically induced apoptosis: Ingrid Druwea, Theresa M. Freudenrich, Kathleen Wallace, Timothy J. Shafer, William R. Mundy. Sensitivity of neuroprogenitor cells to chemical-induced apoptosis using a multiplexed assay suitable for high-throughput screening. doi:10.1016/j.tox.2015.03.011.

Summary: The results demonstrate that, (1) all three commercially available models generated a robust source of proliferating neuroprogenitor cells, and that the assay was sensitive and reproducible when used in a multi-well plate format; (2) there were differences in the response of the rodent and human neuroprogenitor cells to a set of chemicals previously shown to induce apoptosis in vitro; and (3) proliferating neuroprogenitor cells were more sensitive to chemical-induced apoptosis than differentiated neurons, suggesting that neuroprogenitor cells are one of the cell models that should be considered for use in a developmental neurotoxicity screening battery.

Here're more publications of referencing use of hNP1 Neural Progenitors:

Yuji Kaneko, Hideki Shojo, Jack Burns, Meaghan Staples, Naoki Tajiri, Cesar V. Borlongan, DJ-1 ameliorates ischemic cell death in vitro possibly via mitochondrial pathway, Neurobiology of Disease, Available online 21 September 2013, ISSN 0969-9961, http://dx.doi.org/10.1016/j.nbd.2013.09.007...Cell culture and oxygen-glucose deprivation (OGD) hNPCs were obtained from Neuromics...

Xiufang Guo, Severo Spradling, Maria Stancescu, Stephen Lambert, James J. Hickman. Derivation of sensory neurons and neural crest stem cells from human neural progenitor hNP1. Biomaterials, In Press, Corrected Proof,Mar 2013.doi:10.1016/j.biomaterials.2013.02.061 ...hNP1, were obtained from Neuromics (Edina, Minnesota)...

Questions? Do not hesitate to contact me, Pete Shuster: pshuster@neuromics.com or direct phone: 612-801-1007.

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.

The Importance of in vivo Like Astroglial-Neuron Co-Cultures

What We Have Learned

I have many posting on both Neuron and Astroglial Cultures. We have now evolved our solution set to include in vivo like co-cultures. These Astroglial Neuron Co-cultures are designed to mimic in vivo like behaviors. They are potent, pure and proven to work in our clients' unique neurodegenerative disease and toxicity drug discovery assays.

Why is this important? The mix of Astrocytes, Glia and Neurons in your co-cultures can impact your data endpoints and lead to inaccurate conclusions. Here're examples as to how much your data can fluctuate. These are data generated from toxicity assays.

We stand ready to serve you and your team. Questions? Don not hesitate to call 612-801-1007 or e-mail me pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics.

hN2 Primary Human Neurons and Toxicity Assays

Neuromics' hN2 Cells Continue to Shine

I previously posted our Human hN2 Neurons being used for studying the mechanisms of  Nerve Agent VX: hN2 Human Neurons for Toxicity Screening. The versatility of these neurons enabled the researchers to perform a microarray study in which cultured human neural cells were exposed to 0.1 or 10 μM of VX for 1 h. Global gene expression changes were analyzed 6, 24, and 72 h post exposure. Solid primary cell based assay results start with healthy and well behaving cells.

I am pleased to announce success in exposing the cells to a specific cytotoxicity inducing compound (small molecule): 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.

Image: Cytotoxic agent used in dose-response curve

Figure: Dose response curves for 3 specialized somatic cell lines (MRC-5, human primary neurons and human neonatal cardiomyocytes) treated with JC011.

This curve illustrates the sensitivity of human neurons to toxic agents. 

We plan on continuing to use these and our hNP1 Human Neural Progenitors in kinetic, "in vivo like" assays. These assays will give quantitative data on both growth and differentiation inducing agents as well as specifics on how the cells behave when exposed to toxic agents. I will be posting results here.

This data should be of interested to neuro-disease/disorders basic and drug discovery researchers. We plan on making the assays available to researchers. We currently also do small molecule testing and gene expression analysis studies a CRO offering. To learn more, I can be reached at pshuster@neuromics.com or 612-801-1007.

hN2 Human Neurons for Toxicity Screening

Our Human hN2 Neurons proving excellent platforms for Neurotoxicology Studies. I would like to share recent publication and that confirms the potential of these solutions for use in your toxicology assays.

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.

Abstract: Organophosphorus (OP) compounds represent an important group of chemical warfare nerve agents that remains a significant and constant military and civilian threat. OP compounds are considered acting primarily via cholinergic pathways by binding irreversibly to acetylcholinesterase, an important regulator of the neurotransmitter acetylcholine. Many studies over the past years have suggested that other mechanisms of OP toxicity exist, which need to be unraveled by a comprehensive and systematic approach such as genome-wide gene expression analysis. Here we performed a microarray study in which cultured human neural cells were exposed to 0.1 or 10 μM of VX for 1 h. Global gene expression changes were analyzed 6, 24, and 72 h post exposure. Functional annotation and pathway analysis of the differentially expressed genes has revealed many genes, networks and canonical pathways that are related to nervous system development and function, or to neurodegenerative diseases such as Alzheimer’s disease, Huntington’s disease, and Parkinson’s disease. In particular, the neuregulin pathway impacted by VX exposure has important implications in many nervous system diseases including schizophrenia. These results provide useful information valuable in developing suitable antidotes for more effective prevention and treatment of, as well as in developing biomarkers for, VX-induced chronic neurotoxicity.

Images: hN2 Neurons at 18, hrs, 72 hrs and 6 Days.

Image: hN2 cells grown in culture for 4 days and stained with our chicken polyclonal to Neurofilament light or low molecular weight chain NF-L, a marker of neurons. Many of the differentiating cells show strong cytoplasmic and clearly fibrillar staining for NF-L. Blue stain is DAPI and reveals cell nuclei of some non neuronal cells in this culture

I will continue to post updates on new applications of these potent, pure and easy to grow human neurons.

FLICA™ in vitro Caspase Kits for Apoptosis Assays

Great solution for measuring neurotoxicity

Here's a recent pub confirming the capabilities of our FLICA™ in vitro Caspase Apoptosis Detection Kits: Minghai Zhou, Gregory Ottenberg, Gian Franco Sferrazza, and Corinne Ida Lasmézas Highly neurotoxic monomeric α-helical prion protein. PNAS 2012 109 (8) 3113-3118; published ahead of print February 7, 2012, doi:10.1073/pnas.1118090109. ...PFA and stained for caspases using FLICA polycaspase assay (Neuromics) according to the manufacturer's instructions. Caspases...assayed on live cells using the caspase- specific FLICA assay (Neuromics). Cells were photographed using a Nikon inverted epifluorescence...

Images: FLICA was used to assess cell death in primay rat hippocampal neurons.Cells were plated on 25-mm poly-l-lysine-coated coverslips at 300,000 cells per coverslip. Cells were used at 4 or 8 days in vitro. Composite imagae (A) 3 out of 4 cells are apoptotic (green). No cells were necrotic as both of the PI-positive cells were FLICA-positive; they had compromised membranes and were probably in the late stages of apoptosis rather than necrosis. (B) 3 Caspase-positive cells fluoresce green.

I would like to underscore that we have solutions to cover most your needs for Apoptosis and Cytoxicity Assays. I will continue to post updates on customer data and publications.

Cannabinoid Type-1 Receptors and Chemotherapy Related Pain

Chemotherapy can induce painful peripheral neuropathy and also have a toxic effect on peripheral nerves. The authors here show that that cisplatin produces hyperalgesia and toxicity to sensory neurons as indicated by neurochemical, morphological, and functional measures. Increasing AEA signaling at CB1 receptors not only reduced the hyperalgesia but reduced the neurotoxicity of cisplatin as well:  Iryna A. Khasabova,Sergey Khasabov, Justin Paz, Catherine Harding-Rose, Donald A. Simone, and Virginia S. Seybold. Cannabinoid Type-1 Receptor Reduces Pain and Neurotoxicity Produced by Chemotherapy. The Journal of Neuroscience, 16 May 2012, 32(20): 7091-7101; doi: 10.1523/​JNEUROSCI.0403-12.2012.

The authors use our guinea pig TRPV1 antibody to measure cisplatin hyperalgesia vs treated and control mice.

Images: URB597 attenuated effects of cisplatin on protein expression in DRGs. A, TRPV1- and ATF-3-ir were detected by immunofluorescence in L3–L5 DRGs from mice treated with vehicle, cisplatin, or cisplatin plus URB597. Cisplatin (1 mg/kg of body weight, daily for 7 d, i.p.) increased the occurrence of TRPV1- and ATF3-ir in neurons. Co-injection of URB597 (0.3 mg/kg daily, i.p.) with cisplatin attenuated the effect cisplatin on protein-ir. Scale bars: 10 μm (for images within each antigen). B, Quantitative summary of the effect of treatments on TRPV1-ir in neurons. Data are expressed as the mean ± SEM. aSignificantly different from each other group (p < 0.05, one-way ANOVA with Student–Newman–Keuls test; n = 4 mice/treatment). C, Quantitative summary of the effect of treatments on ATF-ir in neurons. Data are expressed as the median and 25th and 75th percentile range. *Significantly different from vehicle control and cisplatin plus URB597 groups (n = 6 mice/treatment; p < 0.001, Kruskal–Wallis ANOVA on ranks test).

Related Reagents:

VR1 N-Terminus (TRPV1)
VR1 (TRPV1)-Goat
VR1 C-Terminus (TRPV1) - mouse specific
All TRP Antibodies
Pain and Inflammation Research Antibodies
Neurotransmission -Neurotransmission Research Antibody Categories

Dr. Steve Stice to Present the Power of StemEZ Neural Cells

Dr. Steve Stice to Present the Power of StemEZ Neural Cells

Posted using ShareThis

I have profiled Steve Stice's research here. The focus has been the excellent research results he and his team at ArunA Biomedical have generated with STEMEZ(TM) hN2 Human Neurons and hNP1 Human Neural Progenitors.

The story continues. He will be presenting the latest at the 9th Annual World Pharmaceutical Congress in Philadelphia, June 14. Topics include: using these neural cell lines to study neurotoxicity in cell-based assays and disease modeling. Recent work conducted in outside laboratories demonstrates that these lines are more sensitive to environmental toxicants than traditional cellular models.

Sample high throughput assay applications:

• Cell morphology and neurite outgrowth


• Cell signaling and transcription factor expression


• Receptor and ion channel function


• Cytotoxicity
  
• Apoptosis, genotoxicity and DNA damage
  

These capabilities has been confirmed by our customers. I look for the use of the STEMEZ cell lines to continue to grow as researchers discover their value in Drug Discovery and Basic Neuroscience capabilities.

Neurotoxicity Testing

We combine our expertise in providing fresh and healthy:

E18 and E20 Rat Primary Neuronal Tissue -NEURON CULTURES

E18 Rat Primary Neuronal Tissue - ASTROCYTE CULTURES

E18 Mouse Neuronal Tissue -NEURON CULTURES

E18 Mouse Neuronal Tissue -ASTROCYTE CULTURES

AND Apoptosis Research Reagents

to help researchers more effectively study Neurotoxicity.

Images: Polycaspase Assay Kit, green was used to assess cell death in primay rat hippocampal neurons.Cells were plated on 25-mm poly-l-lysine-coated coverslips at 300,000 cells per coverslip. Cells were used at 4 or 8 days in vitro. Composite imagae (A) 3 out of 4 cells are apoptotic (green). No cells were necrotic as both of the PI-positive cells were FLICA-positive; they had compromised membranes and were probably in the late stages of apoptosis rather than necrosis. (B) 3 Caspase-positive cells fluoresce green.