Hi
I have be studying on bone derived MSC and in the begining I did passaging, ICC, freezing cells and defrosting cells. There was no problem but now my MSC cell cultures were infected two times and I could not find the reason.
I have suspected;
1. Last time I forgot to wash my cells before trypsinization but this affects only number of cell that I gained.
2. I have realized I was doing differentiation assay at the same time and cells in diffrentiation media were not affected from infection. In my diffrentiation media I was using gentamycine and in control media I used penicillin/streptomycin (Pen/Strep). Is this the reason or Pen/Strep does not work?
3.I recieved the cells on 6/12/2023 and I passaged them 16/12/2023 and I fed them on 19/12/2023. There was no problem and I would feed and passage them on 22/12/2023 but I have learnt they were infected?
I was checking them every feeding time is that enough or should I check them every day under microscope ? I do not want them they are infected or disturbed.
I will be so happy if you share your thoughts because I do not want to lose my cells because of infection third time?
Thank you
Dear Fatma,
In my opinion, contamination occured after feeding at 19-12-2023.
The possible reason:
The complete medium that you used was contaminared, may from be one of more components in making the complete mefium. My suggestion: check all components of complete mefiun. Also, when you feed, add complete nedium via a syringe filter, to assure sterility.
Other possibilities:
1. Problems in your aseptic technic. My suggestion: improve your technic.
2. Contaminants of your culture were penstrep resistant, or penstrep expired and did not work. You can change to another antibiotics. However, if your technic is OK, and all components are sterile, antibiotic is not needed.
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