Neuromics Microglia in Action

 Researchers Study Migration of Neuromics Microglia in Bio-Scaffolds

Check out the publication-https://www.sciencedirect.com/science/article/pii/S2405844023005595.

Images: Migration of Microglia

Here's a link to Neuromics' portfolio of cells for Neurocientists-https://www.neuromics.com/neurons-astrocytes-schwann-cells-microglia.

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.

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.

Human Brain Microvascular Endothelial Cells and BBB

Fc-saxatilin used to Modulate Blood-Brain Barrier (BBB)
Highlights
•Fc-saxatilin prevents VEGF-induced permeability in Neuromics' human brain microvascular cells (HBMECs).
 •Fc-saxatilin inhibits VEGF-induced Src and Fak phosphorylation in HBMECs.
 •Fc-saxatilin blocks the downregulation of claudin-5 expression by VEGF in HBMECS.

Hyun-Jung Choi, Na-Eun Kim, Il Kwon, Dukhwan Choi, Jayoung Kim, Ji Hoe Heo. (2019). Fc-saxatilin inhibits VEGF-induced permeability by regulating claudin-5 expression in human brain microvascular endothelial cells. Microvascular Research. DOI: https://doi.org/10.1016/j.mvr.2019.103953

Figure: Fc–saxatilin attenuates the phosphorylation of Fak induced by VEGF. A) HBMECs were treated with pretreated with vehicle (PBS, 0) or Fc-saxatilin (50, 100, or 300 ng/ml) in the basal medium (0.5% FBS) for 10 min and treated with VEGF (100 ng/ml) for 1 h. Control cells were incubated in the control medium without Fc-saxatilin and VEGF. Cell lysates were subjected to an immunoblot analysis with antibodies against either phospho-Fak, Fak, or actin, as indicated. Representative data from three separate experiments are shown. B) Bands in the immunoblots were quantified using ImageJ and normalized to actin (n = 3); *P < 0.05 compared to the vehicle only-treated control or VEGF-treated control.

These cells are also used to formulate our 3-D Blood-Brain Barrier Model.

Primary Human Brain Pericytes

Pure and Potent
The authors of a chapter on pericytes published in SpringerLink referred to our human primary pericytes as "pericyte-like", but chose not to characterize them because of the "exorbitant cost". Dore-Duffy P., Esen N. (2018) The Microvascular Pericyte: Approaches to Isolation, Characterization, and Cultivation. In: Birbrair A. (eds) Pericyte Biology - Novel Concepts. Advances in Experimental Medicine and Biology, vol 1109. Springer, Cham

Given that these pericytes are also part of our hot selling 3-D Human BBB Model, we are in the process of challenging these claims. We will always take inaccurate claims regarding our solutions seriously, and we take immediate action.

• Characterization of cells-we validated several key markers by immunofluorescence.

Staining of Desmin (dilution 1:100). Secondary antibody conjugated to Alexa 594 (red) and counterstained with DAPI (blue). Cells were mounted using iBrite mounting media.

Staining of Actin (dilution 25 ug/mL). Secondary antibody conjugated to Alexa 594 (red) and counterstained with DAPI (blue). Cells were mounted using iBrite mounting media.

We also plan on doing a phenotypic analysis of these cells and will post here when completed.

• Cost of cells-isolating pericytes from human donors, and expanding to the required number of cells involves much time and effort. We know! Further differentiating stem cells into pericytes is hard and time-consuming. Against this backdrop, we consider our pricing to be inexpensive. Though they are more difficult to derive, they are priced equivalently with our other human primary cells ($789/500,000 USD Cells).

As always, we will post new data here.