Table 1.
Animal age (weeks) | Harvesting | Initial cell density | Plate | Culture medium | Serum | Medium replacement | |
---|---|---|---|---|---|---|---|
Protocol 1 | 6 | Maceration | ∼8 × 107cells | 25cm2 flask | DMEM | 10% and 20% FBS | 7 days |
Protocol 2 | 6 to 8 | Flush | 1 × 107cells | 25cm2 flask | RPMI 2µM L-glutamine 1 × 10−6mol/L hydrocortisone | 10% and 20% HS | 7 days |
Protocol 3 | 6 to 8 | Flush | 2 × 107cells | 25cm2 flask | RPMI 2µM L-glutamine 1 × 10−6mol/L hydrocortisone | 20% HS | 7 days |
Protocol 4 | 16 | Flush | 1.75 × 107cells | 6-well plate | DMEM | 10% FBS | 72 hours |
Protocol 5 | 8 to 10 | Flush and centrifugation | ∼8 × 107cells | 175cm2 flask | RPMI - IMDM 12µM L-glutamine | 9% FBS 9% HS | 24 hours |
DMEM: Dulbecco's Modified Eagle Medium; RPMI-1640: Roswell Park Memorial Institute; IMDM: Iscove's modified Dulbecco's Medium; FBS: Fetal Bovine Serum; HS: horse serum.