I am doing a time-course study on neutrophil apoptosis in cell cultures . So far we have been able to isolate the neutrophils from peripheral human blood with a high yield and viability . However between 0-6 hours of culture we can see white aggregates in the culture that disappears after the 6 hour time-point.
At first we thought it could be contamination of the cell cultures. However, this was disproved after microscopic examination of the cell cultures. Our current suggestion is that it's the neutrophils that are secreting extracellular traps and proteins from their granules after they have been put in culture.
Do anybody have an explanation to what it could be? And how to avoid getting these white aggregates?
Hi,
The white aggregates are sticking to the neutrophils but floating when staining the neutrophils for apoptosis, thereby also falling out of the tubes during the washing steps. Therefore, we lose quite a lot of cells during these steps.
The cells are dying during the culture as they should though.
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