We stain our fibroblast culture (designated for stem cell feeder cells) with AP, the results show a few red stained cells? is this a AP+ results or a false positive? is this means that there is a small stem cell population in our fibroblast culture?
pic attached
sincerely
Indra
I know one thing that AP is an osteogenesis marker when I do differentiate MSCs. Therefore it should not be a marker for MSCs at least.
Sir, AP is a marker and not the stain. Can you please tell which stain did you use? And secondly, the presence of stem cells would be dependant on the source of your fibroblasts. If that's a commercial cell line, the chances are almost nil but if you have isolated that from some mesenchymal origin tissue (mostly adipose, cartilage and bone) you might encounter some stem cells. But again, the chance depends upon the protocol used.
Most importantly, AP will be encountered if you have differentiated your MSCs into osteoblasts. AP has been one of the earliest established osteoblast marker.
its fast red and naphtol stain...and its human adult dermal fibroblast primary isolated, standard culture with DMEM and 10% serum although we do find small population of CD105 and SSEA3 positif cells
what is the chance this being an artefact? And how to rule out this?
I used the ImmPACT Vector Red kit for AP staining and got very bright red signal for stem cells but not feeders(as the pic)...but recently I found that it's not working and still trying to figure out why. Maybe you can try this kit. Also you can observe under microscope for stem cell colonies(should be round and glow on the edge)...
thank you Peggy..i will consider this kit...hope you solve your problem too
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