Dear everyone
I'm working on the endometrium of the uterus. I need to extract endometrial cells from the endometrial tissue. Usually, I will separate the tissue into 2,3 mm pieces and incubated in DMEM/F12 10% FBS with 1% Penicilline/Streptomycin + collagenase 1 mg/ml for 1 hour. After that I centrifuge 1 min at 400x and then remove the pallet, keep the supernatant.
The supernatant will be centrifuged in 5 min at 3000x, then remove the supernatant and keep the pellet. Wash 3 time more with PBS and then seed into 3-cm plate.
But they are continuously contaminated with rod-shaped bacteria, may be Bactobacillus or E coli.
Could I ask if anyone has experience in this topic? could you share with me your experience?
Hello!
I don't have experience on that specific topic, but have you checked the reagents for contamination?
Also, since you are already using ATB (unless those bacteria are resistant), I'd try a deep cleaning of the centrifuge.
María Belén Eyheramonho I have check all, medium, collagenase... incubated for 72 hours but nothing contaminated. So I think the problem is from the tissue, as endometrial tissue always has a microbiota occupying it.
I see. What about antibiotic sensitivity testing? And then using the specific ATB for incubation?
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