Table 1 Seeding, plating, and passaging ratios for primary cell isolation. Isolation components for each cell type include the resuspension volume, plating volume, and number of vessels that can be seeded for primary astrocytes, pericytes, and endothelial cells based on the size of the desired culture vessel. The number of brains used, age of donor, and passage information for each cell type are also included
From: Isolation methods and characterization of primary rat neurovascular cells
Astrocyte & Glia isolation | Â | For one P5-P7 brain | |
Vessel size | Resuspension volume | Plating volume | Number of vessels seeded |
T-75 Flask | 15 mL | 15 mL | 1 |
T-25 Flask | 15 mL | 5 mL | 3 |
35Â mm dish | 15 mL | 1.66 mL | 9 |
Passage astrocytes & glia at 5 DIV in a 1:10 split ratio | |||
Pericyte isolation | Â | For two P56-63 brains | |
Vessel size | Resuspension volume | Plating volume | Number of vessels seeded |
T-75 Flask | 9 mL | 9 mL | 1 |
35Â mm dish | 9 mL | 1 mL | 9 |
Passage pericytes at 15 DIV in a 1:3 split ratio | |||
Endothelial cell isolation | Â | For two P56-63 brains | |
Vessel size | Resuspension volume | Plating volume | Number of vessels seeded |
100Â mm dish | 18 mL | 9 mL | 2 |
35Â mm dish | 18 mL | 1 mL | 18 |
Passage endothelial cells at 5 DIV in a 1:2 split ratio | |||