keratinocytes by flow
mezz001@pn.itnet.it
Mon, 21 Jul 1997 12:12:20 +0200 (MET DST)
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