hi,
is there a rough guide or from your experience, an estimation of cell density being translated to cell confluency? i need this rough translation for initial seeding density.
Hi Iwan this is a tough question as it depends upon your cell line (larger cells have higher % confluence at the same density as smaller cells).
As a rough guide seeding MSC or fibroblasts (quite large cells in 2D) at 10 000 cells/cm2 the cells will be at ~40-50% confluence the next day and ~70-80 % confluence ~ 3 days post seeding.
Please note this is a very rough idea and should be empirically determined for your cell type.
Also as well as fixed cell density you might want to consider fixing the media volume/cm2 across all your flasks or plates too. For example use 0.3 ml per cm2 in both plates and dishes (this helps counteract effect of oxygen effects between different depths of media).
I hope this helps,
Gary
Hi Iwan this is a tough question as it depends upon your cell line (larger cells have higher % confluence at the same density as smaller cells).
As a rough guide seeding MSC or fibroblasts (quite large cells in 2D) at 10 000 cells/cm2 the cells will be at ~40-50% confluence the next day and ~70-80 % confluence ~ 3 days post seeding.
Please note this is a very rough idea and should be empirically determined for your cell type.
Also as well as fixed cell density you might want to consider fixing the media volume/cm2 across all your flasks or plates too. For example use 0.3 ml per cm2 in both plates and dishes (this helps counteract effect of oxygen effects between different depths of media).
I hope this helps,
Gary
Hi Iwan,
Let me present to you translations you've asked for (ThermoFisher).
Set 1. DISHES
Dishes Surface Area (cm2) Seeding Density Cells at Confluency
35mm 9 0.3 x 106 1.2 x 106
60mm 21 0.8 x 106 3.2 x 106 100mm 55 2.2 x 106 8.8 x 106
150mm 152 5.0 x 106 20.0 x 106
Set 2. CULTURE PLATES
6-well 9 0.3 x 106 1.2 x 106
12-well 4 0.1 x 106 0.4 x 106
24-well 2 0.05 x 106 0.2 x 106
Set 3. FLASKS
T-25 25 0.7 x 106 2.8 x 106
T-75 75 2.1 x 106 8.4 x 106
T-160 162 4.6 x 106 18.4 x 106
Best wishes,
Ilya
- Badges
- Science method