I am doing an internship in a cell culture lab and culturing fibroblasts in MEM media supplemented with the appropriate growth factors. After incubation, the media appears healthy: reddish-orange, transparent, and free of turbidity. While observing the culture under a microscope, I identified both adherent fibroblast cells and some suspended cells. However, I also noticed two unidentified structures in the culture flask (as shown in pictures). These structures are [one has sharp edges, and one is ovel in shape.]. I am interested in understanding what these structures could possibly be.
Hello Bilal Rehman
I feel it is debris from FBS. It is usually the result of normal serum components. Fibrin appears in small quantities as fine flake or may occur as a lump of a semi-solid substance. The presence of this material does not alter the serum’s performance and may be mistaken for microbial contamination.
To avoid these in culture, you may allow the serum to stand. The sediments will eventually fall to the bottom of the bottle, and the serum may then be decanted.
Best.
Malcolm Nobre
Dear Malcolm Nobre,
To get rid of the debris from FBS would it be nice to perform centrifugation of the FBS in a 15ml falcom tube? If yes what should be the time and speed that I should consider? I needed to know about this, thank you so much.
Regards,
Bilal Rehman
Do you filter your FBS? If not, consider doing it.
Make sure if the debris is also in the same focus range and whether it moves when you shake the media since particles on top of the culture lid or dust over the lens can also show up. If you're sure it's in the media, filtering the components might be the solution. You can take only PBS/media in a culture plate or an empty plate and check under the microscope to troubleshoot.
I guess, as stated by others that you have no contamination but simply debris. You can routinely filter your medium (containing % FBS) before the culture.
I have answered a similar question to this previously on ResearchGate.
The answer is that these are most likely residual artefacts from the manufacturing process of your TC vessels. As also suggested on this site, they could also be derived from your media formulation (e.g. FBS), be they from an ingredient/component, or contaminant residuals within the vessel you used to prepare your media.
Even when using virgin TC plasticware, such artefacts do occur. Fortunately they are usually sterile, inert, and rarely affect your culture.
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