I cultured Human Umbilical Vein Endothelial Cells (HUVECs) on a plate coated with collagen type 1 (rat tail) at a concentration of 7.5ug/cm^2. The coating procedure involved dilution with cell culture water, incubation for 1 hour at 37°C, followed by removing the solution and washing twice with PBS. Subsequently, I used EGM-2 medium for culturing the cells. Everything appeared normal on the first day, but by the third day, the media became cloudy, and most of the cells were either detached or not visible. Please refer to the attached video for more details. Any insights or suggestions would be greatly appreciated.
I can’t say for sure because you did not provide a view through a phase microscope, but from your description, it is likely your culture is infected with bacteria.
You have bacterial contamination, most likely streptococcus. As you can see in your video, it usually produces the central cloud when you move the dish. Improve your sterility and handling. I also strongly advise using flasks with the filter rather than petri dishes.
Your culture is infected!
Most likely acquired during manual handling and harvest.
Discard. Do not attempt to rescue.
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