Thursday, September 20, 2018

Human Brain Microvascular Endothelial Cell in Action

Recent Pubs
Our Human Brain Microvascular Endothelial Cells (HBMECs) are known for the performance in drug discovery and tox assays. These 21-CFR compliant cells are used in the "blood side" of our 3-D BBB Models.

  1. Shavali Shaik, Bridget Kennis, Shinji Maegawa, Keri Schadler, Yang Yanwen, Keri Callegari, Rishi R. Lulla, Stewart Goldman, Javad Nazarian, Veena Rajaram, Jason Fangusaro, and Vidya Gopalakrishnan. (2018). REST upregulates gremlin to modulate diffuse intrinsic pontine glioma vasculature. Oncotarget. 2018 Jan 12; 9(4): 5233–5250. doi: 10.18632/oncotarget.23750 
  2. Hu et al. (2016). Epigenetic Activation of WNT5A Drives Glioblastoma Stem Cell Differentiation and Invasive Growth. Cell. 167, 1281–1295.,
GREM-1 is required for tube formation in vitro (A) Q-RT-PCR analysis of GREM-1 gene expression in SU-DIPG-IV cells stably expressing either control shRNA or GREM-1-specific shRNA. Lentiviral constructs expressing two different GREM-1 shRNAs (GREM-1.1 and GREM-1.2) were used to knockdown GREM-1. Efficiency of GREM-1 knockdown was determined by Q-RT-PCR and expression was normalized to 18s RNA. Significance is as shown (**less than.01). (B-C) HUVEC or human brain microvascular endothelial cells (HBMEC) were cultured in endothelial cell medium and or conditioned medium from either control shRNA or shGREM-1.2 transfected SU-DIPG-IV cells. Tube formation in matrigel was measured after 16h and images were obtained. The ability of GREM-1 to rescue loss of tube formation upon REST knockdown was determined by addition of human-recombinant GREM-1 (rGREM-1) to conditioned media-endothelial media mix. Scale bars, 100μm. (C) Quantification of tubes in matrigel shown in Figure B (right panels). Data shown is mean +/- SD, ***p less than .001, n=3. (D) Western blot analysis to assess VEGFR2 levels in SU-DIPG-IV, -VI and –XIII cells, HUVECs and HBMECs was done using anti-VEGFR2 antibodies. Tubulin served as a loading control. (E) Western blot analysis was performed to assess AKT signaling downstream of GREM-1 interaction with its potential receptor VEGFR2 in HUVEC and HBMEC. Anti-pAKT (S473), anti-pAKT (T308), total AKT, and anti-actin were employed.
If these cells fit your assay requirements and need more data/info, do not hesitate to contact me, Pete Shuster, CEO at or cell: 612-801-1007.

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