We are using small molecules to induce iPS differentiation.
The first photo show alkaline phosphatase screening for control iPS and after treating iPS with small molecule.
2nd and 3rd photos showing some iPS differentiate colonies after introducing the small molecules.
My question is how we could detect the differentiated cell while we don't know the types of obtained cells?
Does anyone have any suggestions?
In photo No.2, we can see a little number of iPS cells and increasing in cells size, how we could explain?
I think it is difficult to detect all types of differentiated cells. I only can think about the immunostaining with the antibody specific to most common cell types in the ES cell differentiation (ie. SMA).
You could try a PCR array that detect markers of the different differentiation pathwyays, like this kit:
http://www.qiagen.com/products/catalog/assay-technologies/real-time-pcr-and-rt-pcr-reagents/rt2-profiler-pcr-arrays?catno=PAHS-048Z#geneglobe
Few cells have morphologies that can allow to recognize them (neuronal cells, fibroblasts) but for the majority this is simply not possible. So immuno-staining and PCR can give a hint to what cells are obtained after differentiation. I would only suggest that the selection of the markers to check for should also take into account the current knowledge of the small molecules used: which pathways do they activate or inhibit? which lineages are dependent on these pathways during embryonic development in vivo?
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