Neural Progenitors and Cool Science

Titania Nanotubes and Neural Prostheses

I am always on the hunt for the use of our Neural Progenitor Markers in cool and important human health applications. Here our Nestin Antibody is use to access the growth and maintenance of C17.2 neural stem cell line cultured in these nanotubes: Jonathan A. Sorkin, Stephen Hughes, Paulo Soares, Ketul C. Popat. Titania nanotube arrays as interfaces for neural prostheses. Materials Science and Engineering: C Volume 49, 1 April 2015, Pages 735–745. doi:10.1016/j.msec.2015.01.077

Image: Nestin staining eSC Derived hNP1 Human Neural Progenitors. Cells were stained using goat anti-mouse Alexa Fluor 488 secondary antibody (green) (Molecular Probe, A-11001) and counterstained with DAPI (blue).

Abstract: Neural prostheses have become ever more acceptable treatments for many different types of neurological damage and disease. Here we investigate the use of two different morphologies of titania nanotube arrays as interfaces to advance the longevity and effectiveness of these prostheses. The nanotube arrays were characterized for their nanotopography, crystallinity, conductivity, wettability, surface mechanical properties and adsorption of key proteins: fibrinogen, albumin and laminin. The loosely packed nanotube arrays fabricated using a diethylene glycol based electrolyte, contained a higher presence of the anatase crystal phase and were subsequently more conductive. These arrays yielded surfaces with higher wettability and lower modulus than the densely packed nanotube arrays fabricated using water based electrolyte. Further the adhesion, proliferation and differentiation of the C17.2 neural stem cell line was investigated on the nanotube arrays. The proliferation ratio of the cells as well as the level of neuronal differentiation was seen to increase on the loosely packed arrays. The results indicate that loosely packed nanotube arrays similar to the ones produced here with a DEG based electrolyte, may provide a favorable template for growth and maintenance of C17.2 neural stem cell line.

I will continue to how our stem cell solutions are used in cool, new discoveries.

Sensitive, Specific and Cost Effective Protein Expression Analysis

Reaching for Cell Based Assay Solutions Excellence

We have been receiving positive feedback from clients using our Quantibody® and RayBio® C-Series Membrane-Based Antibody Arrays. Our manufacturing partner, Raybiotech, is ISO 9000 certified insuring all arrays are made to exacting specification. The cost for measuring a single cytokine runs as low as 16 USD/Cytokine!

We use these arrays to test the blood serum of clients with autoimmune diseases to determine levels of cytokine and growth factor expression vs healthy controls. We also use them to determine proteins secreted by our  USB Derived hMSCs cultured in our hMSC Media. This gives us insight as to the protein expression patterns in our hMSC, Chondrogenesis and Osteogenesis Assays.

Figure: The graph shows cytokine secretion of hMSCs after a continuous 24 day culture. These results were obtained using our select markers from our cytokine and bone metabolism arrays. Data courtesy of Dr. Jim Musick and Tiana Tonrey, Vitro Biopharma.

We are pleased to announce the addition of these new human and mouse arrays:

Quantibody® Human Growth Factor+Neurotrophin+Cytokine Array Q1-10. Precisely measures: Amphiregulin, BDNF, bFGF, BMP-4, BMP-5, BMP-7, beta-NGF, EGF, EGFR, EG-VEGF (PK1), FGF-4,FGF-7 (KGF), GDF-15, GDNF, Growth Hormone, HB-EGF, HGF, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-6, IGF-1, Insulin, M-CSF R, NGFR (TNFRSF16), NT-3, NT-4, Osteoprotegerin (TNFRSF11B), PDGF-AA, PLGF, SCF, SCF R (CD117/c-kit), TGF alpha, TGF beta 1, TGF beta 3, VEGF-A, VEGFR2, VEGFR3 and VEGF-D.
Mouse C-Series Cytokine Array. C1-2-Semi-quantitavely measures: GCSF,GM-CSF, IL-2, IL-3, IL-4, IL-5, IL-6, IL-6, IL-9, IL-10, IL-12 p40/p70, IL-12 p70, IL-13, IL-17A, IFN-gamma, MCP-1 (CCL2), MCP-5, RANTES (CCL5), SCF, TNF RI (TNFRSF1A), TNF alpha, Thrombopoietin, (TPO) and VEGF-A.

Images: Representative images obtained with RayBio® C-Series Antibody Arrays. These membranes were probed with conditioned media from two different cell lines. Membranes were exposed with Kodak X-Omat® film at room temperature for 1 minute. Note the strong signals of the Positive Control spots in the upper left and lower right corners.

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

Human Growth Factor+Neurotrophin+Cytokine Array-Test 40 Markers in on Assay

Sensitive, Specific and Cost Effective

Neuro-immuno and degenerative diseases and disorders commonly show dysregulation of Growth Factors and Cytokines. Using our Quantibody Neuroscience Arrays, we have measured the blood serum of clients with Neuro-inflammatory/immune response diseases/disorders including Autism Spectrum Disorder (ASD). Most showed lowered levels of Growth Factors/Neurotrophins and elevated levels of Inflammatory Response Cytokines.

In order to expand the number of biomarkers measured, we are pleased to announce the addition of our New Growth Factor+Neurotrophin+Cytokine Array. This will enable us to further determine the "finger prints" of these diseases/disorders at the protein level. This array includes: Immunogen: Amphiregulin, BDNF, bFGF, BMP-4, BMP-5, BMP-7, beta-NGF, EGF, EGFR, EG-VEGF (PK1), FGF-4,FGF-7 (KGF), GDF-15, GDNF, Growth Hormone, HB-EGF, HGF, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IGFBP-6, IGF-1, Insulin, M-CSF R, NGFR (TNFRSF16), NT-3, NT-4, Osteoprotegerin (TNFRSF11B), PDGF-AA, PLGF, SCF, SCF R (CD117/c-kit), TGF alpha, TGF beta 1, TGF beta 3, VEGF-A, VEGFR2, VEGFR3 and VEGF-D.

I will continue to post testing results here.

Solutions for Studying Neuro-degeneration

Data Rich and Frequently Published

The Neuromics' brand is built, in part, by our proven ability to provide solutions for the study of neuro-degeneration. These include:

• Neuronal-Glial Markers
• Primary Neurons and Astrocytes
• Apoptosis Research Reagents
• Transfection Reagents

A recent example shows the use of one of  MAP-2 markers to study hearing decline with age: Radtke-schuller S, Seeler S and Grothe B(2015) Restricted loss of olivocochlear but not vestibular efferent neurons in the senescent gerbil (Meriones unguiculatus). Front. Aging Neurosci. 7:4. doi:10.3389/fnagi.2015.00004.

Figure: Lipofuscin granules in MSO neurons of an aged gerbil. MSO neurons are MAP2 immunostained (Alexa Fluor 647, red). Lipofuscin granules have been excited with the DAPI excitation wavelengths and appear blue. Confocal images show a maximum projection of image stacks in A and a single optical image of 0.3 µm thickness in the enlargement in B. Scale bar in A: 50 µm and 20 µm in B.

We stand ready to serve you. Should you have interest or questions, do not hesitate to contact me directly: Pete Shuster-Owner/CEO-pshuster@neuromics.com or direct phone: 612-801-1007. Thank you.

Cancer Associated Fibroblasts (CAFs)-Flow Cytometry and Immunostaining Results

If they Walk and Talk Like CAFs, They Are!

We are experiencing growing demand for are CAFs. They are closely associated with primary tumor cells and participate in the neoplastic process. There is reciprocal communication between CAFs and tumor cells through paracrine effects of secreted growth factors, cytokines & chemokines from both fibroblasts, tumor cells and other tumor-associated cells.

Potential users are asking how well the cells are characterized. The purpose of this posting is to share key gene expression data:

CELL LINES	CD105	CD90	CD44	CD326	CD133	FAP
SC00A1 (hMSCs)	+	+	+	-	-	-
SC00A5 (pancreatic fibroblasts)	+	+	+	-	-	-
CAF05 (colorectal CAFs)	+	+	+	-	-	+
CAF07-AD (adenocarcinoma lung CAFs)	+	+	+	-	-	+
CAF08 (Pancreatic Stellate CAFs)	+	+	+	-	-	+

Table: expression of key proteins

Images: Human Lung Adenocarcinoma CAFs stained with Vimentin, aSMA, b-Catenin and DAPI (blue),

Cancer Associated Fibroblasts Gene Expression Data(pdf - 784Kb)-Flow Cytometry and Immunostaining Results.

Questions? Please contact me directly-Pete Shuster- CEO & Owner-pshuster@neuromics.com or 612-801-1007.

Racing into 2015

Thank you for a Great 2014!
Wow. I would like to thank our customers, collaborators and friends for helping us achieve unprecedented success in 2014.

We plan on racing hard into 2015. I would like to highlight our focus areas for the New Year.

Stem Cell Activating Solutions-We have been pilot testing solutions foir balanced immunity and stem cell vitality. Our clients have been mostly peopole with autoimmune related issues. After 6 months of therapy, the improvement in symptoms and test results, for many, has been stunning (see below).
We have developed migration and other assays to test these solutions in vitro using our UCB Derived hMesenchymal Stem Cells. We also use medical testing to prove and improve their therapeutic value. These assays and arrays are available for labs and clinics interested in testing stem cell related solutions. Here're representative data from our Quantibody Array testing program.


Stem and Primary Cell Based Assays-We will continue to add new solutions for your assays. These will include mixed culture that more closely mimic in vivo environments like mixed astroglial-neuron cultures. Below is our hN2^TM Human Neurons stained with one of our neuron markers.

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.

• Here're some of our human cells we added in 2014:
  Motor Neurons-GFP Labeled-Potent and Easy to Culture Mouse Motor Neurons
• Human Endothelial Cells-New-Artery, Microvascular, Vein & Umbilical Cord Derived + Culture Media
• Cancer-Associated Fibroblasts (CAFs) and Cells-Solutions for studying tumor growth, angiogenesis and metastasis.

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Check out and join my Stem Cell Clinical Trails Group on Linkedin (4500+members strong).
We wish you a rewarding 2015.

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Pete Shuster-
CEO & Owner--pshuster@neuromics.com or 612-801-1007

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