Magic Red and Cancer Research

Measure Apoptosis in Real Time in Cancer Cells

Magic Red™ (MR) Kits measure apoptosis via active caspases and cathepsins in whole living, intact cells - no lysis required. As apoptosis progresses and caspase activity increases, the red fluorescent signal increases.

In this example, Magic Red is used to measure apoptosis in HeLa Cells after treatment with Artesunate (ART), an anti-malarial drug and potential chemotherapy agent.

Figure: ART activates lysosomal function. A, HeLa cells were first treated with ART (50 μM) for 12 h, followed by LTR labeling for 30 min, and further analyzed using confocal microscopy. B, HeLa cells were loaded with LTR following time course drug treatment, and fluorescence intensity of 10,000 cells/sample was measured by flow cytometry. C, HeLa cells were treated as indicated in A and then incubated with Magic Red cathepsin L and LTG for 30 min and observed under a confocal microscope. D, HepG2 cells were treated as in A and then labeled with LTR described earlier in A. E, HepG2 cells were treated as in A, followed by a Magic Red cathepsin B and L assay for 45 min. Fluorescence intensity of 10,000 cells/sample was measured using flow cytometry. F, HeLa cells were first loaded with DQ Red BSA for 1 h and then treated with ART (50 μM) for 18 h and incubated with LTG for 30 min. G, HeLa cells were treated with ART for 18 h and then immunostained for LAMP1. H, LAMP1 signals of 80 cells from three independent experiments were quantified by ImageJ (Student's t test, #, no significance). I, HeLa cells were treated with ART for the indicated time, and then Western blot was performed to detect the LAMP1 protein level. Scale bar, 10 μm. Error bars, S.D. doi: 10.1074/jbc.M114.564567

We have a variety of Apoptosis Kits for testing potential Chemotherapeutic Agents.

Pancreatic Cancer Tumor Cell Assays

Providing Pancreatic Tumor Cells or Related Custom Assays

We are pleased to announce the addition of Pancreatic Tumor Cells to our Cell Based Assay offerings. We are also generating a wealth of internal data with our Tumor Cells and Cancer Associated Fibroblasts. We have now opened up our capabilities to Cancer Researchers via our new CRO services. These included customized assays for studying toxicity using compounds/small molecules. Here's an example:

In addition, we can do kinetic, live cell screens using these cells plus our hMSC and CAFs. To learn more call 612 -801-1007 e-mail me: pshuster@neuromics.com. We stand ready to serve you in a way that fits your specific requirements.

Osteoblast Activators and Musculoskeletal Disease Assays

We now have the ability to precisely test human Osteoblast activating agents. Tools available include:

1.  Live cell screening
2. Quantibody Bone Metabolism Arrays
3. PCR
4. Flow Cytometry
5. Immunostaining
6. Western Blotting

This means we can generate data that provides you a clear picture of the effects of small molecules/compounds on the activation, expansion and migration of Osteoblasts. Here's an example.

Image: Activin A expression from Osteoblasts treated with Activators vs Controls

We will soon be releasing a turn-key assay for Osteoporosis Drug Discovery. We can also run custom assays that fit your specific requirements. To learn more, you can call (612-801-1007) or e-mail: pshuster@neuromics.compshuster@neuromics.com). Thank you. 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.

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.