I am culturing Rhesus Monkeys PBMC, using the ficoll isolation method, and trying to expand the cell substitution by adding PHA in IL-2 containing medium. However, a large proportion of bacilliform cells resulted, and it doesn't look like the E.coli since it stays in situ. I uploaded the picture where the arrows depict the bacilliform cell, still I take a cell VCR which you can find that a large ratio of not clear cell substitution.
To exclude the bacteria and the reagent contamination, I did the gram stain first, re-cultured the "medium" without the Rhm PBMC. The results show that there was no bacteria (16sRNA not done yet). Do you have any idea about this observation?
What was the result of the gram stain? I suggest going ahead with the 16S rRNA.
I agree with Dr Hassan, a quick 16S rRNA shoud clarify the situation
I have done the 16sRNA test, no clearly expected band exists. And when observing the cells under the microscope again, these bacilliforom cells disappeared. Is it possible that these ones were swallowed by the lymphocyte?
Dear Dr. Morceau,
I am not clear about its sources and compositions either, however it disappeared a few days later, and it didn't have much effect on cell differentiation, we also really want to know the causality.
Best
Hi Suzy An,
It might be related with the difference of each individual. Have you tried another source of Rhm PBMCs? You may found that some species differentiate better.
We also tried other isolation method, like "Lymphocyte Separation tube" method, there were also bacilliforms . This suggested that these forms may come from the monkey's own blood, I do not know how to explain this phenomenon either. If you have new findings, could you please tell me?
Best
Have you ruled out platelets? Human PBMC extractions always have some platelets present. The size seems consistent with that.
We did not ruled out the platelets, and if you keep culturing for several days, change fresh meidum, there would be less left.
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