Reagent

Volume

Sample (peripheral blood)

100 μl

RPMI (HEPES and glutamine)

5 ml

Fetal Bovine Serum

500 μl

Phytohemagglutinin

10 μl

Streptopenicillin

100 μl

Colchicine

100 μl

Hypotonic solution at 0.56%

Total incubation time of 60 hs

Incubation temperature 28˚C