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Primary cultures of bovine retinal microvascular pericytes (BRPC) are
often isolated from freshly isolated calf eyes obtained from a local abattoir
using homogenization and a series of filtration steps. BRPC are often
subsequently cultured in DMEM with 5.5 mM glucose and 20% FBS (Hyclone
Lab. Inc., South, UT) on fibronectin (NYBen Reagents, New York Blood Center)
coated dishes (Costar, Cambridge, MA). Cells are most commonly cultured
in 5% CO2 at 37°C, media changed every 3 days and cells passaged 1:2 when
confluent. Cells at passages 2 to 7 are usually used for experiments.
Pericyte homogeneity can be characterized for their homogeneity by immunoreactivity
with monoclonal antibody 3G5 and confocal microscopy. |
