i am using A549 cells its growing very fast and i am facing problem in differentiating cells and contamination but it is attending confluence very fast, i am using T25 flask. I can see dead cells, one day after passage
Hey Sunilgowda, cancer cells usually grow very fast. You can get rid of dead cells by changing the media. To ensure contamination, you keep a T25 flask with only media along with your A549 culture. If you see something is growing in the media, then your A549 culture may have contamination. Hope this helps. Good luck.
@rajdeep thank you sir, yes i did that no turbidity in the media thank you for good suggestion
Hello Sunilgowda S N
Dead cells usually end up floating in the medium where as bacterial contamination do not float in the medium.you can easily find out by observing the bacterial movement and morphology that is cocci or rods shaped etc. You can trace back your contamination by placing just the medium / PBS / FBS- FCS in the incubator overnight and replace those that are contaminated
Make sure you have added the necessary antibiotics and antimycotics to the medium before culturing any cells. Also since you are growing cancer cells and you are observing fast growth that leads to subculture the very next day I would suggest you culture them in T 75mL culture flask which gives the cells more space to grow and you could subculture them after few days
@Zubin Zaheer thank you for your good suggestion, i will try with T75 as u said it will make space rather than T25 and other than bacterial contamination and other contaminants wont float?. Thank you once again
Well it does when there is not enough space left for them to grow also it is important to distinguish non-living particulates such as chemical contaminants and contaminating bacteria or fungal. There are two kinds of movements basically- the "Brownian" movement and "Active" movement. cell debris and any non-living particulates exhibit Brownian movement where they exhibit some movement, but pretty much stay in the same place. In the cases of bacterial contamination, you will see active movement where these particles move actively and move away from the current position. Next time, you see particulates, look at the movement patterns :)
Hope this helps.
- Badges
- Science topic