Friday, November 22, 2019

Optimizing Human Microglia Cultures

Healthy and Happy Cells
Microglia, one of the glial cell types in the CNS, is an important integral component of neuroglia cell network. They act as brain macrophages when programmed cell death occurs during brain development or when the CNS is injured or pathologically damaged.

This makes them an important tool for drug discovery.

We have research proven Human Microglia.

Neuromic's Microglia Cultured in T25 Flask
Related Protocol:
  • Place the flask in a clean Biosafety cabinet. 
  • Remove the seal on the T-25 Flask. 
  • Remove about 15-20ml of media from the flask. Change the cap on the flask, make sure media is not touching the cap, close the cap. 
  •  Culture the cells in a 37oC incubator with 5%CO2
  • For best result, do not disturb the culture Flask for at least 72 hours after receiving the T-25 flask the culture has been in. Change the growth medium the next day.
Need Primary Human Cells? Just ask me. pshuster@neuromics.com.

Tuesday, November 12, 2019

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 quanti´Čüed 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.