Description

Lewis Rat Primary Kidney Glomerular Endothelial Cells from Cell Biologics are isolated from aorta of 4-8 week old laboratory Lewis rat. Lewis Rat Primary Kidney Glomerular Endothelial Cells are grown in T75 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Cell Biologics’ Culture Complete Growth Medium generally for 3-7 days. Cultures are then expanded. Prior to shipping, cells at passage 3 are detached from flasks and immediately cryo-preserved in vials. Each vial contains 0.5x10⁶ cells per ml and is delivered frozen. The method we use to isolate primary endothelial cells was developed based on a combination of established and our proprietary methods. These cells are pre-coated with PECAM-1 (CD31) antibody, followed by the application of magnetic beads pre-coated with secondary antibody.

Product Testing

Lewis Rat Primary Kidney Glomerular Endothelial Cells are tested for expression of markers using antibody, PECAM-1 Antibody (M-20, sc-1506, Santa Cruz) or ZO-1 Rabbit Polyclonal Antibody (Catalog No. 617300, Life Technologies) by immunofluorescence staining. Lewis Rat Primary Kidney Glomerular Endothelial Cells are also tested for uptake of Dil-Ac-LDL (Catalog No. L-35353, Invitrogen), a functional marker for endothelial cells. Lewis Rat Primary Kidney Glomerular Endothelial Cells are negative for bacteria, yeast, fungi, and mycoplasma. Cells can be expanded for 3-7 passages at a split ratio of 1:2 or 1:3 under the cell culture conditions specified by Cell Biologics. Repeated freezing and thawing of cells is not recommended.

Suggested Medium

Endothelial Cell Medium /w Kit – 500 ml

Catalog No. M1266

Laboratory Applications

Lewis Rat Primary Kidney Glomerular Endothelial Cells can be used in assays of cell-cell adhesion, migration, vascular tube formation, transendothelial resistance (TER). Standard biochemical procedures performed with cell cultures include RT-PCR, western blotting, immunoprecipitation, immunofluorescent staining, immunofluorescent flow cytometry, or generating cell derivatives for desired research applications.

Storage

Cell Biologics ships frozen cells on dry ice. On receipt, immediately transfer frozen cells to liquid nitrogen

(-180 °C) until ready for experimental use. Live cell shipment is also available on request.

Never can primary cells be kept at -20 °C.

Documents

RA-6014G.LS

Cell-Culture-Protocol-CB

Disclaimer

Investigators should handle the cells that they receive from Cell Biologics with caution and treat all animal cells as potential pathogens, since no test procedure can completely guarantee the absence of infectious agents.