I purchased some Hi5 cells from Invitrogen which I grew up and then adapted to suspension. I froze the adapted cells at varying densities (as the literature is varied), between 0.5 × 10^7 - 1 x 10^7 viable cells/vial. The manufacturers guidelines state that cells frozen at higher densities may be thawed directly into suspension conditions using the appropriate seeding density. But I cannot locate any literature that gives a protocol or a seeding density.

Does anyone have any suggestions as to what seeding density should be used to initiate the adapted Hi5 cells directly into suspension?