Study of endothelial cell growth, differentiation and influence of oncogenes.

Mouse SV129 brain endothelioma

The cell line b.End3 has been established from brain endothelial cells of SV129 mice. Immortalisation has been carried out by infection of primary cells with retrovirus coding for the Polyoma virus middle T-antigen. b.End3 cells are positive for endothelial specific proteins (PECAM-1, Endoglin, MECA-32, Flk-1) tested by FACS. Inflammatory cytokines induce the expression of proteins such as ICAM-1, VCAM-1 and E-selectin. Injection into mice causes development of haemangiomas.

DMEM + 2mM Glutamine + 5μM 2-Mercaptoethanol (2ME) + 1mM Sodium Pyruvate (NaP) + 1% Non Essential Amino Acids (NEAA) + 10% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:4 i.e. seeding at 4x10,000 cells/cm² using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37°C.

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