Floating of adherent cells during culture is a common but also a headache.
The phenomenon of a large number of adherent cells floating can lead to serious consequences such as cell loss and inaccurate experimental results. Why do large numbers of cells float? And how can it be remedied? How can this be prevented? In this issue, we will learn about cell culture with D1 Biotech.
cell culture
Possible reasons for the large number of cells floating
and solutions
The quality of the experimental reagents is poor
In vitro culture experiments of cells, fetal bovine serum is often added to provide nutritional support for cells. The usual addition concentration is between 5-20%, accounting for a large proportion. If the quality of fetal bovine serum is poor, it may cause cells to die due to insufficient nutrients, and a large number of cells will float.
Solution: Select experimental reagents such as fetal bovine serum with guaranteed quality, and conduct pre-experiments in a small range before officially starting the experiment to verify the effect of the experimental reagents.
Contamination of the cell culture system
If the cell culture system is accidentally contaminated with bacteria, fungi, and mycoplasma, a large area of cells may float.
Solution: Discard the contaminated cells and clean up the experimental environment with a contamination removal reagent, such as MyCoplasma OFF from MB in Germany.
The cell density is too high
When the cell density is too high, competition between cells increases, causing some cells to float away from the substrate.
Solution: Perform regular cell passaging to avoid excessive cell density. Depending on the cell type, an appropriate range of densities can generally be found in the literature.
Poor cell quality
Poor-quality cells may have poor adhesion and difficulty attaching to the surface of the dish.
Solution: Ensure that you use high-quality media and reagents, regularly check cell morphology and growth status, and avoid using aged cells.
Improper surface treatment of the Petri dish
If the coating or treatment on the surface of the dish is uneven or improper, it can cause the cells to not attach evenly.
Solution: Use pre-treated Petri dishes, or choose the appropriate coating (e.g., collagen, gelatin, poly-L-lysine, etc.) according to the needs of the experiment.
Incubation conditions are not suitable
Inappropriate conditions such as culture temperature, CO2 concentration, and humidity affected the adhesion and growth of cells.
Solution: Adjust the appropriate culture conditions, such as temperature, humidity, CO2 concentration, etc., according to the cell type and culture requirements.
Cellular aging
If the cell culture time is too long, the cells may age and lose their ability to adhere.
Solution: Replace fresh cell lines regularly and control the incubation time of cells.