Rat Bone Marrow-Derived Endothelial Cells from Creative Bioarray are isolated from bone marrow tissue of 6-8 week old laboratory Sprague-Dawley rat. Rat Bone Marrow-Derived Endothelial Cells are grown in T75 tissue culture flasks pre-coated with gelatin-based coating solution for 2 min and incubated in Creative Bioarray’ 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 at least 1x10^6 cells per ml and is delivered frozen. The method we use to isolate endothelial cells was developed based on a combination of established and our proprietary methods. These cells are pre-coated with PECAM-1 (CD31) antibody, following the application of magnetic beads pre-coated with secondary antibody.
Rat Bone Marrow-Derived 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. Rat Bone Marrow-Derived Endothelial Cells are also tested for uptake of Dil-Ac-LDL (Catalog No. L-35353, Invitrogen), a functional marker for endothelial cells. Rat Bone Marrow-Derived 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 Creative Bioarray. Repeated freezing and thawing of cells is not recommended.
Storage and Shipping
Creative Bioarray 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.
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