Subculture Protocol for A549 human alveolar adenocarcinoma cell line

A549 (ATCC number CCL-185)A549 is a human epithelial cell line derived from a lung carcinoma tissue the cells are adherent in culture.

Subculture Protocol for A549 human alveolar adenocarcinoma cell line

1.     Remove the media, rinse cells with HANKS solution

2.     Add 3 mL of Trypsin-EDTA to flask and watch for cell layer dispersal under an inverted microscope. This should occur within 1-1.5 minutesand watch for cells  dispersal under inverted microscope.

3.     Remove Trypsin-EDTA ansd wait for 3-5 mintues.

4.     Add 2-4 mL of growth medium to cells with pipette, gentlymixed and remove 75-90% of the cells.

5.     Add 8-10 mL of growth medium to cells with pipette , incubate at 37°C in 5% CO2.

6.     Split confluent culture 1:4 to 1:9 every 4-7 days with trypsin/EDTA. Make sure to renew medium every 2-3 days.