I'm Culturing adult mice cardiomyoctes and during changing medium step the cells seems washed away. Any tips about how to avoid loosing cells during washing step?
We make the following experience every day in the cell culture laboratory:
In order not to lose the cells during washing,
1: The cells should be vital,
2: The bottle or Petri dish should be subconfluent (60-70% of the area should be occupied by the cells),
3: The washing medium (preferably PBS) should not be very cold,
4: The nutrient medium should be adequately equipped with the additives,
5: Wash procedure should be done gently and neatly,
6: The culture bottle, Petri dish, PBS, nutrient medium and incubator should not be contaminated,
7: Incubator should be well equipped with temperature and Co2
Hello dear Youjing Zheng ,
You should coat the flask or dish with proper substances like laminin.
Hi
When changing the culture medium, consider a few points: 1- Be sure of the appropriate temperature (37) of the culture medium. 2- You should keep the plate tilted to prevent the tip tip from hitting the cells so that the old culture medium flows to one side and it is easier to remove it. 3- When adding fresh medium, do not pour it directly on the cells, but gently pour it from the plate wall. 4 - As a general recommendation: to increase cells adhesion, you can add a higher percentage of serum to the culture medium, for example, 15-20% instead of 10%. Good luck.
We make the following experience every day in the cell culture laboratory:
In order not to lose the cells during washing,
1: The cells should be vital,
2: The bottle or Petri dish should be subconfluent (60-70% of the area should be occupied by the cells),
3: The washing medium (preferably PBS) should not be very cold,
4: The nutrient medium should be adequately equipped with the additives,
5: Wash procedure should be done gently and neatly,
6: The culture bottle, Petri dish, PBS, nutrient medium and incubator should not be contaminated,
7: Incubator should be well equipped with temperature and Co2