primary fibroblast culture,
I am doing a primary culture of fibroblasts from skin explants, which confluence is considered to make the pass
1. confluence under the microscope
2. the confluence that the entire well of the 6 plate is full
Thank you!
Hello Maria Fernanda Rosado
When the cells reach 80% confluence, they must be sub cultured to ensure proper growth and health of the cells. 80% confluency of cells means 80% of the surface of the well should be covered with cells.
You would have to observe the cells under the microscope for 80% confluency. The second option would result in 100% confluence or over confluency which is usually not recommended while subculturing the cells.
Best.
Usually 80-85% confluency is considered optimum for passaging when seen through microscope. Always use lower and higher magnification for clear picture.
80% is a good starting point for passaging primary cells. You can check the confluency with low magnification microscope. Best
thanks to everyone!
Malcolm Nobre
Vikas Vitthal Andhale
Benjamin Fournier
The confluence of a primary fibroblast culture should be determined by microscopic observation. The cells should be near 100% confluent, with no visible gaps between them. The entire well of a 6-well plate being full is not a reliable indicator of confluence, as the cells may not be densely packed enough for passaging.
If primary fibroblasts are allowed to grow to confluency, they will become contacted inhibited, which will make passaging them difficult. Subculture when 75-80% confluent.
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