I have been beating my head for an explanation on what this generic term really means. Does it refer to the contents of a cell post lysis? Does it refer to the fluid collected after pelleting a cell culture? Or is it the media/fluid in which a cell is suspended during growth?
Dear Jonas, I think the meaning of 'cell culture supernatant' is fairly easily defined if you don't want to think too far - it is simply the fluid (and suspended solids including some cells) after centrifugation under particular centrifugal force (xg) for a specified time. Usually if you spin your sample fast enough you can be reasonably sure that whole cells and large cell debris are removed in the pellet - but this is never absolute, especially if you just pour the supernatant off the pellet. I would suggest that a 'lysed cell supernatant' would be a better phrase to refer to the centrifuged solution following cell lysis. The term 'supernatant' is probably best restricted to the fluid that remains after centrifugation, and not be used to describe the fluid component of a mixture before centrifugation. Regards, Andrew
The term "supernatant" is mainly refers to the media/fluid in which cells are suspended/cultured during growth.
Dear Jonas, I think the meaning of 'cell culture supernatant' is fairly easily defined if you don't want to think too far - it is simply the fluid (and suspended solids including some cells) after centrifugation under particular centrifugal force (xg) for a specified time. Usually if you spin your sample fast enough you can be reasonably sure that whole cells and large cell debris are removed in the pellet - but this is never absolute, especially if you just pour the supernatant off the pellet. I would suggest that a 'lysed cell supernatant' would be a better phrase to refer to the centrifuged solution following cell lysis. The term 'supernatant' is probably best restricted to the fluid that remains after centrifugation, and not be used to describe the fluid component of a mixture before centrifugation. Regards, Andrew
Am following this with interest. Just to confirm, is it the broth that is to be centrifuged and then the supernatant taken? What exactly are we centrifuging???
To clarify, David, the cell culture supernatant would be obtained by centrifugation of the cell, suspended in their culture medium, under the centrifugation conditions specified. as Andrew Spiers stated, the composition of the resultant supernatant will depend on the speed (and therefore G-force) and duration of the centrifugation, and the nature of the cells and other components in the culture medium, including whether lysed or not.
Good afternoon, why is it that in the supernatant medium of the cell monolayer there are dark structures with no defined shape? What are they and why are they formed?
cell free broth or fluid collected after precipitation of bacterial cells by centrifugation
The word supernatant means swimming fluid. It can be either media that cell suspended in during growth or media collected after centrification.
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