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.

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.

Extracellular Matrix Environment and Chemotherapeutics

Substrate Matters!

Neuromics' has been promoting a variety of 3-D Cell Based Assay Solutions for the past several years. These include: Nanofibers, Hydrogels and Extracellular Matrix (ECM) Proteins. We have found that the adoption rate for these as standard tools for drug discovery is slower that we anticipated.

We believe substrate matters so I am pleased to share a recent publication that references use of our Collagen IV and other ECM proteins. This confirms the importance of using a more in vivo like environment in testing chemotherapeutics: Thuy V. Nguyen,Marianne Sleiman,Timothy Moriarty,William G. Herrick,Shelly R. Peyton. Sorafenib resistance and JNK signaling in carcinoma during extracellular matrix stiffening. Publication: Biomaterials. Elsevier. 13 April 2014. http://dx.doi.org/10.1016/j.biomaterials.2014.03.058.

Abstract: Tumor progression is coincident with mechanochemical changes in the extracellular matrix (ECM). We hypothesized that tumor stroma stiffening, alongside a shift in the ECM composition from a basement membrane-like microenvironment toward a dense network of collagen-rich fibers during tumorigenesis, confers resistance to otherwise powerful chemotherapeutics. To test this hypothesis, we created a high-throughput drug screening platform based on our poly(ethylene glycol)-phosphorylcholine (PEG-PC) hydrogel system, and customized it to capture the stiffness and integrin-binding profile of in vivo tumors. We report that the efficacy of a Raf kinase inhibitor, sorafenib, is reduced on stiff, collagen-rich microenvironments, independent of ROCK activity. Instead, sustained activation of JNK mediated this resistance, and combining a JNK inhibitor with sorafenib eliminated stiffness-mediated resistance in triple negative breast cancer cells. Surprisingly, neither ERK nor p38 appears to mediate sorafenib resistance, and instead, either ERK or p38 inhibition rescued sorafenib resistance during JNK inhibition, suggesting negative crosstalk between these signaling pathways on stiff, collagen-rich environments. Overall, we discovered that β1 integrin and its downstream effector JNK mediate sorafenib resistance during tumor stiffening. These results also highlight the need for more advanced cell culture platforms, such as our high-throughput PEG-PC system, with which to screen chemotherapeutics.

Figure: High-throughput biomaterial platform for drug screening. (A) The high-throughput platform consists of a black-walled, glass bottom plate, with PEG-PC gels cast in each of the inner 6x10 wells. (B) Gels can be functionalized with any protein or peptide of interest, and they support the adhesion and growth of carcinoma cells. We used this platform to test carcinoma cell response to a kinase inhibitor (sorafenib) as a function of underlying gel stiffness and ECM adhesive protein cocktail. (C) A representative graph of SkBr3 proliferation (y-axis) in response to sorafenib (x-axis) across a range of gel stiffness (colors) demonstrates the IC-50 calculation.

This confirms the importance of considering your substrate environment when developing your in vitro assays for High Content and High Throughput Drug Discovery. We will continue to provide updates.

Increasing Returns in Drug Discovery

"Harnessing the Power of Cells"
 

Cell based assays presentation v1_03_2012 from Pete Shuster

This presentation provides an executive overview of how our Solutions can improve Drug Discovery processes.

Why Consider our Solutions?

Research proven solutions used by large Pharmas, Biotechs, Academic and Government Labs. 2500+ customers-Publications/Testimonials
Lower per well costs-we can aggressively discount cells and media
Customer success focused                                  
Expert technical support
Detailed protocols and methods
Quick replacements and reorders
Large expert resource network
Published expert input
Non confidential; customer feedback and data sharing through social media outlets and blogs