I have cultured RASF from synovial fluid of rheumatoid arthritis patients. Even after two passages I can see a lot of granules surrounding the fibroblast in the culture flask, I want to know if these are lymphocytes/ macrophages or is it some contamination? While taking the picture on the computer it seems to be moving so I am worried if its a bacterial contamination. If it remains in the flask then the expression profile would be different from that of pure fibroblast.
Sir, please find the picture in the attachment. I am interested to know about the dots on fibroblasts and in the media. I had added anti Mycoplasma and antibiotics. I found similar pictures in many of the published papers on RASF but i am curious to know that why it is not clean and after how many passages i can expect it to have fibroblasts only. This picture is of 2nd passage from the synovial fluid of RA patient. The 2nd picture is of OA patient from the 2nd passage, it also looks very similar to that of the FLS from RA patient. Is there any morphological difference between the FLS of RA and OA??
I would absolutely agree with Max. I would add just a quick point about nomenclature. Fibroblast-like synoviocte (FLS) is an outdated term for a fibroblast in the intima of synovium in a tissue section, or otherwised hypothesised to be in the intima. In culture all you know is that you have fibroblasts of synovial origin, or synovial fibroblasts. There is no guarantee that they will be from the intima. Intimal cells have quite different properties from deeper synovial fibroblasts, although there is some evidence for all synovial fibroblasts having a specialised repertoire of gene expression response to stimuli. There is a vast literature using the term FLS for cultured cells for no good reason - it is best avoided.
Synovial fibroblasts from RA and OA sources have been found to show different patterns of gene expression that may persist through subculture but there is no real way of knowing what this means or whether it is an effect of sampling.
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