Dear flowers, has anybody been thinking of (or has anybody set up) a way to recognize differentiated vs undifferentiated keratinocytes by flow? When these cells differentiate they form the so called "cornified envelope" which is an actual envelope of keratins that, in the end, wraps up the cell. A test to verify the number of keratinocytes that have formed the envelope in a primary culture is already available in the literature and is based on cell counting followed by lysis with an SDS-containing buffer and subsequent counting of the remaing unsoluble envelopes. The % envelopes out of the total cell number prior to the lysis represents an indication of the differentiation of the culture. I am interested in setting up the assay using flow cytometry and think that it should not be an impossible thing to do. Essentially, I shoud be able to sort out keratinocytes with and without the envelope, and maybe this could be done using an antibody against some protein of the envelope. Has anybody ever done something like that or is anybody willing to give some, very much appreciated, suggestion about it? Thank you very much for your help Daniela Karrer Drug Discovery Dept. Istituto di Ricerca C. Serono Rome (Italy) tel 39-6-919491 fax 39-6-91949203 e-mail : mezz001@pn.itnet.it
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