hi
i need a second opinion as to whether my podocyte cell lines are differentiating. these are SVI podocytes and they proliferate at 33 degrees celcius, differentiate at 38 degrees celcius. It requires at least 14 days to differentiate.
podocytes at proliferative temperature exhibit cobblestone morphology while at differentiated state exhibit arborized morphology like dendrites. I am not sure (inexperienced) if there is differentiation taking place, as from online sources the differentiated podocytes really look arborized. mine may look a bit arborized but nowhere near as arborized.
attached are the images of my podocytes at different day point of 38 degrees.
images from top to bottom: day 6 differentiation (first 2 images), day 13 differentiation (3rd and 4th image), undifferentiated state (5th image)
Given that WT-1 expression was restricted to podocyte nuclei in normal adult kidney, you should perform F-IHC to confirm the differentiation.
http://image.search.yahoo.co.jp/search?rkf=2&ei=UTF-8&gdr=1&p=podocyte+differentiation+marker#mode%3Ddetail%26index%3D9%26st%3D0
To confirm whether your cells have differentiated you will have to check the expression of differentiation markers of the podocytes e.g. nephrin, podocin, CD2AP, and synaptopodin, by immunofluorescence or western blotting.
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