Neuromics' Microvascular Endothelial Cells in Action

 Widely Used and Data Rich

Neuromics Human Brain Microvascular Endothelial Cells (HBMECS) are frequently used and data rich. Here are HBMECS in culture showing early microtubular formation.

HBMECS in culture showing early microtubular formation

They are an excellent component for building Blood-Brain Barrier (BBB) Models. Questions? E-mail Mr. David Rupp-david@neuromics.com. 

Cancer Associated Fibroblasts and Cell Lines

 Wide Variety

Neuromics has a growing catalog of CAFS and Cell Lines. Here's an example of our Breast CAFS.

Breast CAFs in culture. Grown using Stem Cell Complete Low Serum Media (cat. CAFM02)

Cells are prepared and manufactured in an ISO 9001:2015 certified environment and ISO 5 cleanroom.

Questions? email david@neuromics.com

Neuromics' Human Neurons in Action

Our primary human neurons (cat. HNC001) have been busy lately, continuing to become one of our most popular and versatile products. Earlier this month, Japanese researchers published their findings when creating 3D brain-like spheroids containing our primary human neurons. Their results suggest the spheroids mimic in vivo conditions and are suitable for use as an in vitro neurological disease model. Check out the publication for yourself here.

Human Neurons in Culture

Since the beginning of 2022, our neurons have been cited a number of times spanning various research areas. They were used by researchers to suggest a link between the COVID-19 virus and Alzheimer's disease (lean more here). Additionally, investigators published findings (see here) using the neurons to find a potential therapeutics target for neuroblastoma.

Questions? Do not hesitate to contact me pshuster@neuromics.com or 612-801-1007. Pete Shuster-CEO and Owner

Potent and Pure Fetal Bovine Serum

Potent and pure FBS. Single use, triple filtered and manufactured under the strictest guidelines, Pricing starts at $245/500 ml

Check us out Fetal Bovine Serum.

Introducing our Prostate Cancer Cells Lines

 Check out the Options

Neuromics is pleased to introduce many of the top human prostate cancer cell lines to our already expansive selection of cell offerings. To complement the DU 145 cells we added earlier this year, Neuromics is now offering VcaP, LNCaP, and PC-3 cells to complete our catalog of human prostate cancer cell lines.

Image: VcaP cells in culture.

VCaP cells are known for their high androgen receptor and PSA expression. Unlike most other prostate cancer cell lines, LNCaP cells are highly sensitive to androgens and have a low tumorigenicity. Meanwhile, PC-3 cells are noted for their high metastatic potential along with their insensitivity to androgens and fibroblast growth factors. Lastly, DU 145 cells are not hormone-sensitive and do not express PSA.

Neuromics' Human Brian Microvascular Endothelial Cells in Action

Fc-saxatilin inhibits VEGF-induced permeability by regulating claudin-5 expression in human brain microvascular endothelial cells (HBMVECs)

Neuromics' cells were at the heart of this study.

Highlights: 

Fc-saxatilin prevents VEGF-induced permeability in human brain microvascular cells (HBMVECs). 

Fc-saxatilin inhibits VEGF-induced Src and Fak phosphorylation in HBMVECs. 

Fc-saxatilin blocks the downregulation of claudin-5 expression by VEGF in HBMVECs.

Image: HMBECS in Culture

Protocol: Human brain microvascular endothelial cells (HBMVECs) were purchased from Neuromics (HEC02; Minneapolis, MN, USA). HBMVECs were maintained in ENDO-growth medium (MED001; Neuromics). 

Here's a link to the full study: https://www.sciencedirect.com/science/article/pii/S0026286219301177#!

TRPV1 and Migraines

New Links to Insulin

Hungarian researchers used our guinea pig polyclonal TRPV1 receptor antibody (GP14100) to help answer why heightened insulin levels can lead to migraines.

Immunohistochemistry in rat trigeminal ganglion. (A) Trigeminal ganglion neurons in the ophthalmic division immunoreactive for insulin receptor (InsR), TRPV1 receptor and CGRP. (B) Coexpression of insulin receptor (InsR) with TRPV1 receptor and/or CGRP in trigeminal ganglion neurons. Colours representing insulin receptor-, TRPV1 receptor- and CGRP-immunoreactivities and their coexpressions indicated in B applies also to A. (C) A trigeminal ganglion neuron retrogradely labeled with True blue expresses both insulin receptor (InsR) and TRPV1 receptor

Conclusion-the present findings indicate that insulin may activate TRPV1 receptors in the trigeminovascular system. Modified TRPV1 receptor function induced by insulin may also increase the sensitivity of both neural and vascular TRPV1 receptor for its agonists. Our data may provide a pathophysiological basis for the increased incidence of migraine in patients with hyperinsulin levels.

About IBA Lifesciences

IBA Lifesciences' proprietary Strep-tag® technology exploits one of the strongest non-covalent interactions in nature: the interaction of biotin and streptavidin. The system is based on the highly selective and easily controllable interaction between the synthetic Strep-tag®II peptide and the specially engineered streptavidin, called Strep-Tactin®, which is one of the most stable proteins known. The Strep-tag®II binds specifically to the engineered streptavidins, Strep-Tactin® and Strep-Tactin®XT, by occupying the binding pocket of the natural ligand biotin. Hence, the interaction is easily reversible by excessive addition of the competitor.

Strep-tag®II consists of eight amino acids (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys), whereas the Twin-Strep-tag® includes this motif two times in series connected by a linker and is accordingly composed of 28 amino acids. Both exhibit intrinsic, although unequal, affinity towards the streptavidin derivative Strep-Tactin® and its successor Strep-Tactin®XT: The binding affinity of Strep-tag®II to Strep-Tactin® (1µM) is nearly 100 times higher than to streptavidin. A further improvement was achieved by the development of Strep-Tactin®XT, which shares a nM affinity with the Strep-tag®II and a pM affinity with the Twin-Strep-tag®.

To Learn more watch this intro video/

Studying Diabetic Retinopathy?

Check out How Neuromic's Tools Are Used

The authors used four of Neuromics to complete this study-

Sulodexide reduces glucose induced senescence in human retinal endothelial cells A. Gericke, K. Suminska-Jasińska & A. Bręborowicz Scientific Reports volume 11, Article number: 11532 (2021) Cite this article.

Material and Methods

Experiments were performed on HREC (#HEC09, Neuromics, Edina, MN, USA) in in vitro culture. Cells were seeded in 75 cm² culture flasks coated with AlphaBioCoat Solution (#AC00, Neuromics, Edina, MN, USA) and were grown in Endo Growth Medium (EKG001, Neuromics, Edina, MN, USA) supplemented with fetal bovine serum 0.5% until they formed monolayers. Then the cells were harvested with cell detachment solution (#ADF001, Neuromics, Edina, MN, USA) and seeded in quadruplicates into 25 cm² coated, as described above, culture flasks at a density of 1.5 × 10⁵ cells/flask.

We conclude that Sulodexide may have a beneficial effect in cases of diabetic retinopathy. It slows down hyperglycemia-dependent senescence of endothelial cells, which translates into the lower angiogenic and inflammatory impact of these cells. An important observation was that Sulodexide also has effective antiangiogenic and anti-inflammatory effects in the senescent HREC. That means that Sulodexide may be effective in retinal endothelial cells, which are already senescent. Further studies are required to explain the potential effect of Sulodexide on the endothelial glycocalyx structure and permeability of the retinal endothelial layer composed of the senescent cells to molecules with various size and electrical charge.

We are pleased to see our solutions used in this important  study. It could lead to drugs that slow diabetic retinopathy.

Neuromics' FBS and National Cancer Institutes

 Best Price and Performance 

I was honored to work with Jessey at National Cancer Institute (NCI).

The were determining whether to change partners. According to Jessey, there were 24 lab PIs who would be supplied with the winner's FBS Although we proved competitive, we were not selected.

The study was conducted with only premium grade FBS (most expensive). We are announcing a new Preferred FBS this Monday 10/25. It works well for supplementing define media and works great for most human primary sells. And works fabulous for human and animal cancer cells.

Interested in learning more? Contact me at pshuster@neuromics.com or 612-801-1007. Pete Shuster CEO, Neuromics.

Neuromics-3D Human Blood Brain Barrier!

Compound Penetration Study

  

Classification: Low permeability: Pe < 1.00 nm/s 

Moderate permeability: 1.00 < Pe < 10.0 nm/s 

High permeability: Pe > 10.0 nm/s 

* The signal responses of Atenolol , 

Sulpiride, and Cimetidine in receiver samples were undetectable.

For the convenience of calculation, 

1/300 of the measured peak area ratio (PAR) of the relevant AD (A>B Donor) 

samples were used as the PAR values of Atenolol , Sulpiride, and Cimetidine 

in receiver samples.

Check out the study-

https://www.neuromics.com/ittrium/reference/D8xf65cx8x1/BBB%20Penetration%20Data.pdf  

Neuromics' Renal Proximal Tubule Cells in Action

Human Renal Proximal Tubule Cells Express Klotho

Our human primary cells are widely used and frequently referenced in a variety of publications. In this study, our RPT Cells were used to study the impact of Klothko expression on protection of kidneys in diabetics: Meng Xue, Feng Yang, Ying Le, Yanlin Yang, Bingsen Wang, Yijie Jia, Zongji Zheng, and Yaoming Xue. (2021). Klotho Protects Against Diabetic Kidney Disease via AMPK- and ERK-Mediated Autophagy. Acta Diabetologica, 32. doi: 10.1007/s00592-021-01736-4.

Klotho expression in human renal proximal tubule cells after high-glucose stimulation. (a) Levels of Klotho mRNA and (b) Klotho protein were assessed in human renal proximal tubule cells treated with low glucose and high glu- cose. (c) Morphological observations of Klotho expression by confocal microscopy

Klotho improved renal tubular cell autophagy via the AMPK and ERK path- ways and played a role in renal protection. These findings provide new insight into the mechanism of Klotho and autophagy in DKD.

Trigeminal Neuralgia (Facial Pain)

 Markers for Pain Research

Trigeminal Neuralgia is a progress chronic condition. Dysfunctional trigeminal signaling can lead to intense, searing facial pain. It is caused by pressure on the trigeminal nerve.

The root causes of the condition are not well understood. Our pain research markers have been widely used and frequently published through the years and have been important contributors to our growth, Here's a recent pub referencing use of one our P2X3 Antibodies-Momoko Koizumi, Sayaka Asano, Akihiko Furukawa, Yoshinori Hayashi, Suzuro Hitomi, Ikuko Shibuta, Katsuhiko Hayashi, Fusao Kato, Koichi Iwata, and Masamichi Shinoda. (2021). P2X3 Receptor Upregulation in Trigeminal Ganglion Neurons Through TNFα Production in Macrophages Contributes to Trigeminal Neuropathic Pain in Rats. The Journal of Headache and Pain, 22, 31. doi: 10.1186/s10194-021-01244-4

Images: Changes in P2X3R expression in TG neurons innervating whisker pad skin and the involvement of P2X3R signaling in TG neurons in orofacial mechanical hypersensitivity on day 7 following TNC. a Photomicrograph of FG-labeled P2X3IR TG neurons following the TNC or sham procedure. Arrows indicate FG-labeled P2X3IR TG neurons. Scale bar: 100 μm. b Mean number of FG-labeled TG neurons ipsilateral to the TNC or sham procedure (n = 5 in each). c The frequency of FG-labeled P2X3-IR TG neurons ipsilateral to the TNC or sham procedure. * p < 0.05 vs. sham. (n = 5 in each, student’s t-test). d The time course of changes in the MHWT by subcutaneous A317491 or TNP-ATP administration on day 7 following the TNC or sham procedure.

The signaling of TNFα released from the activated macrophages and infiltrated into and/or proliferated in the TG induces the upregulation of P2X3R expression in TG neurons innervating the orofacial region, resulting in orofacial mechanical allodynia following TNC. Consequently, TG neuronal P2X3R hyperexpression by enhanced TNFα signaling in the TG is a potential target for TN treatment..

Malaria and the Blood-Brain Barrier

 EPCR and ICAM Receptors

Neuromics Astrocytes and Pericytes were used in this important study-Yvonne Adams, Rebecca W. Olsen, Anja Bengtsson, Nanna Dalgaard, Mykola Zdioruk, Sanghamitra Satpathi, Prativa K. Behera, Praveen K. Sahu, Sean E. Lawler, Klaus Qvortrup, Samuel C. Wassmer, and Anja T.R. Jensen. (2021). Plasmodium Falciparum Erythrocyte Membrane Protein 1 Variants Induce Cell Swelling and Disrupt the Blood–Brain Barrier in Cerebral Malaria. Journal of Experimental Medicine, 218 (3). doi: 10.1084/jem.20201266.

This is the first study showing malaria pericytes interacting with human endothelial cells. Please note we have the complete BBB Model that can be used for studies like these.

Human Schwann Cells that Work

 No Doubt!

Sometimes we hear our human cells did not express a certain receptor in the hands of our customers. We guarantee our cell "walk and talk" as advertised. If you are ever in doubt. we will test for expression of any receptors or cytoplasmic proteins. We also do this as part of our quality checks.

Here're, for example, is a pub showing our Schwann Cells working. Note the receptors tested here. Seung Min Shin, Francie Moehring, Brandon Itson-Zoske, Fan Fan, Cheryl L. Stucky, Quinn H. Hogan, and Hongwei Yu. (2021). Piezo2 Mechanosensitive Ion Channel is Located to Sensory Neurons and Non-Neuronal Cells in Rat Peripheral Sensory Pathway: Implications in Pain. bioRxiv. doi: 10.1101/2021.01.20.427483

Figure. IHC delineation of Piezo2 (PZ2) axonal component and Schwann cell expression. Representative montage images on sciatic nerve cryosections show double immunostaining (IS) of Piezo2 (red) with a selection of neuronal markers (green), including Tubb3 (A), NF200 (B), IB4 (C), and CGRP (D). Representative montage images on sciatic nerve cryosections show double- IS of Piezo2 (red) with glia markers (green), including S100 (E), p75NTR (F), GFAP (G), and MBP (H). Representative montage images on human Schwann cells (I) and isolated Schwann cells (J) from rat sciatic nerve show double-IS of Piezo2 (red) with S100 (green). Scale bars: 50 um for all.

Check out our growing catalog of primary human cells today https://www.neuromics.com/human-cells-tissue. Satisfaction guaranteed or your money back.

Cancer Associated Fibroblasts (CAFS) Traction Dynamics and Remodeling in 3D Matrices

 Neuromics' Colorectal CAFS in Action

In a first of its kind study, the mechanical properties of our CAFS were determined. This is important because, understanding how these cell behave in vivo will help improve the efficacy of new therapies: Bashar Emon, Zhengwei Li, Md Saddam Hossain Joy, Umnia Doha, Farhad Kosari, and M Taher A Saif. (2020). A Novel Method for Sensor-Based Quantification of Single/Multi-Cellular Traction Dynamics and Remodeling in 3D Matrices. bioRxiv. doi: 10.1101/2020.09.24.311647

"Cells in vivo generate mechanical forces (traction) on surrounding 3D extra cellular matrix (ECM) and cells. Such traction and biochemical cues may remodel the matrix, e.g. increase stiffness, which in turn influences cell functions and forces. This dynamic reciprocity mediates development and tumorigenesis. Currently, there is no method available to directly quantify single cell traction and matrix remodeling in 3D. Here, we introduce a method to fulfil this long-standing need. We developed a high-resolution microfabricated sensor which hosts a 3D cell-ECM tissue formed by self-assembly. It measures cell forces and tissue-stiffness and can apply mechanical stimulation to the tissue. We measured single and multicellular force dynamics of fibroblasts (3T3), human colon (FET) and lung (A549) cancer cells and cancer associated fibroblasts (CAF05) with 1 nN resolution. Single cells show significant force fluctuations in 3D. FET/CAF co-culture system, mimicking cancer tumor microenvironment, increased tissue stiffness by 3 times within 24 hours."

Here's one of 10 videos illustrating these properties.

 CAFS are widely used and frequently published. Check them out today!

Neuromics' n-Fect Delivers

 Gene Expression Tools in Action

Neuromics has a wide offering of transfection reagents that are perfect for both in vivo and in vitro applications. We take pride in seeing our product used in research, which is certainly true of our transfection reagents (see here).

Furthermore, we'd like to highlight a recent use of our n-Fect Transfection Kit (cat.# NF30150) in a doctoral thesis recently published. The paper looked into the neuroscience of alcohol addiction, looking towards the purinergic family receptor P2X4, which has been linked to alcohol addiction in mouse models. Using our n-Fect Transfection Kit, the molecular and cellular mechanisms linking P2X4 expression and voluntary alcohol consumption are explored. Our n-Fect reagents were used on brain slices containing the ventral tegmental area (VTA) of mice. They conclude that purinergic control of VTA neurons is a neurotransmitter system that should be further studied as a target for alcohol dependence drugs.

Image: Examples of p-Fect (Cat.# PF3000) transfection in different cell types.

Questions? Do not hesitate to contact Rose Ludescher, VP of customer satisfaction -rose@neuromics.com

Premium Virus Transport Media vs PBS

 There is a difference

Here's a video that illustrates the difference between CDC recommended Virus Transport Media and Sterile PBS (salt water). It answers the questions "what is in your VTM" and "why does it matter". Please consider sharing so it can find its way to those in need!

Check out our VTM-https://www.neuromics.com/virus-transport-media

METHOD OF TREATING CENTRAL NERVOUS SYSTEM DISEASE

 Blood-Brain Barrier (BBB) Permeability Assay

Yes, These are troubled times. With our current focus on eradicating COVID-19, we forget that many of the world's most insidious and costly diseases have limited treatments and no cures.

That is why Neuromics continues to develop its human based 3-D Cell-Based Assays. At the center if our offerings is our BBB Model.

Here's a recent patent application that includes a permeability assay using our model-Sookhee Bang, Jeong Kuen Song, Seung-wook Shin, Kwan Hee Lee, and Ho June Lee. (2020). Method of Treating Central Nervous System Disease. United States Patent Application 20200230218

BBB Permeability Assay of AL04 The in vitro human BBB model (Neuromics, USA) was established using co-cultures of primary Human brain endothelial cells (HBEC), Human brain pericytes (HBPC), and Human brain astrocytes (HBAC). In vitro human BBB model kit has two sides (luminal, blood/abluminal, brain) with 12 transwell inserts (polyester membrane, 0.4 um pore, diameter 12 mm, insert growth area: 1.12 cm2). HBPC were grown on the bottom side of the inserts, HBEC were monolayered on the upper side of the inserts, and HBAC were grown on the bottom of the 12-well culture plate (Neuromics, USA). In vitro BBB model was activated according to the manufacturer's instructions for 4 days. Briefly, the medium from both luminal and abluminal (lower, brain) sides of the transwell insert was changed every other day. Before the transport experiment, the abluminal side was filled with the permeability assay medium. For the permeability assay, purified AL04 or recombinant HSA (Sigma, USA) were added to the luminal side (0.3 ml) of the transwell insert to yield a final concentration of 1 or 10 uM. Incubations were performed on orbital shaker (100 rpm) at 37° C. Samples (150 μl) were collected from the abluminal side (1.2 ml) at 60, 120 and 240 min and immediately replaced with fresh permeability assay medium. The concentrations of transported AL04 or rHSA were measured by Human albumin quantitation ELISA kit (Bethyl laboratory, USA) and analyzed using the standard curve method. Permeability coefficients (Pe, cm s−1) were calculated using the equation: Pe=(VA/(A×c0))×(dQ/dt), where VA is volume of assay buffer in blood side (inside of the insert), A is the surface area of the insert (1.12 cm2), c0 the initial concentration of protein sample added into blood side, dQ/dt the concentration of transported protein sample in brain side in a defined time period. The permeability coefficients (Pe, cm s−1) for the purified AL04 or recombinant HSA were calculated as previously described (Prades, 2015; Nakagawa, 2009).

If you have interest and want to learn more, please email Rose Ludescher, VP, Customer Satisfaction-rose@neuromics.com

Be safe; be healthy.

What's In Your Virus Transport Media (VTM)