Description

Mouse Embryonic Cells-derived Endothelial Cells (ES-ECs) from Cell Biologics are isolated from mouse Embryonic fibrolasts of pathogen-free laboratory mice. iPS-ECs are grown in T25 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 are detached from flasks and immediately cryo-preserved in vials. Each vial contains at least 1x10⁶ cells per ml and are 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 antibody, followed by the application of magnetic pre-coated with secondary antibody.

Product Testing

ES-ECs are tested for expression of markers using antibody, VE-cadherin (CD144, VE-cadherin Antibody, C-19, sc6458, Santa Cruz); AF1002 (R&D System) or CD31/PECAM-1 (Purified Rat Anti-Mouse CD31, Catalog No. 553370, BD) by immunofluorescence staining or FACS. C57BL/6 Mouse Primary Aortic Endothelial Cells are negative for bacteria, yeast, fungi and mycoplasma. Cells can be expanded for 3-6 passages at a split ratio of 1:2 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. M1168

Laboratory Applications

ES-ECs can be used in assays of cell-cell adhesion, migration, vascular tube formation, or transendothelial resistance (TER). Standard biochemical procedures performed with endothelial 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. Primary cells can never be kept at -20 °C.

Documents

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.