Question

summery about Thawing Sf9 insect cells

Answer 1

Frozen sf9 cells are stored in liquid N2 in its vapor phase frozen cells are directly supplied in or thawed in sf900 SFM

procedure for thawing

1 Rapidly thaw frozen vial in a 37°C water bath.

2Triturate and transfer the entire contents of the cryovial into a 125 mL shake flask containing 27 mL of pre-warmed Sf-900 II SFM,

3incubate in a 28o C ± 0.5o C non-humidified, ambient air-regulated incubator or warm room on an orbital shaker platform rotating at 125-150 rpm. Loosen caps of flasks to allow oxygenation/aeration.

Once the culture density has reached >2 x 106 viable cells/mL, determine viable and total cell counts .

. Expand Sf9 cultures by seeding shake flasks at 3 to 5 x 105 viable cells/mL by diluting cells in pre-warmed growth medium.

Subculturing Cells to maintain and subculture Sf9 cells in suspension or adherent culture.