A few buddings are observed in my Vero cell culture under microscopy view (both 10x and 40x) and I'm suspecting those are yeast contamination. The vero cells grew relatively slow and required replenishment with complete medium a few times to achieve 80% confluence in T-25 flask.
You are right. The culture is contaminated with yeast. It is better to discard the culture as it is difficult to get rid of yeast contamination.
You are right. The culture is contaminated with yeast. It is better to discard the culture as it is difficult to get rid of yeast contamination.
It is possible to rescue contaminated cell cultures by centrifugation.
Here the protocol:
- put 5ml HISTOPAQUE 1077 in a 15mL falcon.
- carefully pipette single cell suspension of the cell culture on top
- centrifuge 25 min (700x g, no brake!)
- yeast will pellet at the bottom of the tube (yellow- green pellet)
- cells will stay at histopaque /media interface
- take off cell clound and respend in 10 ml media
- centrifuge 4min (700x g) and plate the pelleted cells in new dish.
You might have to do it a couple of times in order to get rid of all the yeast.
Good luck!
Steffen
Thanks Mrinalini Sharma
Indeed, both microscopy view and cell condition suggest that the culture is most likely susceptible to yeast contamination and so it was thrown away to prevent further contamination to other cultures.
Thanks Steffen Storck , ones would definitely try that if the culture is precious for the lab
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