I am culturing rat adipose tissue mesenchymal stem cell in DMEM low glucose 10% FBS media, after 1 or 2 days of culture I am getting this type of debris or contamination(shown in the picture: red circle). Please guide me are these contamination and why are these coming ?
Try adding antibiotics and antifungal (amphotericin b) to culture medium, enzymatic and wash solution. For example: (50mL culture medium)
44 mL DMEM
5 mL FBS
500 uL penicillin
500 uL amphotericin b
I'm not a big fan of combating microbes with antibiotics/antifungal agents (just use aseptic technique). Do you see turbidity and pH changes (if using phenol red)? Do all your cultures have this or is it just this one?
Can Kiessling There is no visible turbidity or media color change. I am getting these things in all cultured cells but these cells are showing in higher amount.
Jahaira Cortez I am already using antibiotic antimycotic solution(A002 Himedia) in the culture medium.
Not to rule out contamination, this could also be FBS protein clumps. Did you heat inactivate as per
https://www.researchgate.net/post/Can-anyone-tell-me-if-my-fetal-bovine-serum-is-contaminated-see-attached-pictures
?
One way to monitor contamination, just take a sample of the media, add to a flask/plate (6, 12, 24 whatever) and just leave it there for a few days. Monitor the pH, turbidity and phase contrast observation. You can also DAPI stain to see if you have nuclear DNA staining. If your cells are adherent cells, you shouldn't really have (many) DAPI positive stains when you take a sample of the medium. There will be some floaters obviously.
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