Dr. Jessica Wagener, Application Specialist Cell Handling at EppendorfThis article appeared first in “Inside Cell Culture”, the monthly newsletter for cell culture professionals.
- The impact of standardized cell seeding protocol
- The time factor and cell sedimentation
- Video: Correct pipetting technique
- Air bubble formation
- Video: How to avoid air bubble formation in cell seeding
- Homogeneous cell adhesion
- Cell density dependent behavior
Why is cell seeding a critical step in all cell culture experiments?
Cell seeding is usually the first protocol step and a standard procedure in cell-based experiments. A correct and standardized cell seeding protocol is a critical factor for reproducible experimental results. The main challenge in this step is to achieve and maintain comparable cell numbers in all repeated experiments. Variations in seeding cell numbers and the formation of air bubbles during seeding will increase standard deviations, making your results less reliable. Therefore, several factors need to be considered when establishing a solid cell seeding protocol in your lab.
The time factor in cell seeding protocols
When you seed your cells, from a 15 mL tube into a multi-well plate, for example, it will take some time until you have filled all the wells. No matter if you use a single-channel or a multi-channel pipette in your cell seeding protocol, the longer the process takes, the more cells will sediment in the tube. So, without mixing the cell suspension in the tube or reservoir, you will get varying cell numbers from one well to another (Image 1).