[實驗技術]細胞培養面積

以下為常用細胞培養皿粗略培養面積

100 mm dish ＝ 58.1 cm2   1.2 x 10^5
60 mm dish = 20.8 cm2
35 mm dish =9.6 cm2

6 well- plate = 9.6 cm2 (4x 10^4)
12 well- plate = 3.8 cm2 (2x 10^ 4)
24 well- plate = 1.8 cm2  (10^4)
48 well- plate = 0.75 cm2

3t3-L1 cell 培養條件 (From ATCC culture method )

Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Never allow culture to become completely confluent.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
   The recommended inoculum is 2 to 3 X 103 cells/cm2. Subculture before cultures become 70 to 80% confluent or when cells reach 5 to 6 X104 viable cells/cm2.
6. Incubate cultures at 37°C.

Interval: Every three days

Medium Renewal: 2 to 3 times per week