Annexin V/PI assays for adherent cell lines.

From: Michael Ormerod (Michael_Ormerod@compuserve.com)
Date: Tue May 02 2000 - 04:24:06 EST


After incubation of ovarian carcinoma cell lines with cis-platin, the
apopttoic cells detach from the culture dish and can be found flowting in
the sulture medium (for example, see: Cisplatin induces apoptosis in a
human ovarian carcinoma cell line without concomitant internucleosomal
degradation of DNA.  M.G. Ormerod, C.F. O'Neill, D. Robertson and K.R.
Harrap. Expl. Cell Res.,206, 231-237 (1994);
Cis-diamminedichloroplatinum(II) induced cell death through apoptosis in
sensitive and resistant human ovarian carcinoma cell lines.  M. G. Ormerod,
C. F. O'Neill, D. Robertson, L. Kelland and K. R. Harrap. Cancer
Chemotherapy and Pharmacology, 37, 463-471, 1996).

In my experience, there are few apoptotic cells to be fuond still attached
to the culture dish.

To count the number of apoptotic cells, having confirmed microscopically
that the 'floating' cells are indeed apoptotic, you only need to count the
number of detached cells.

Michael Ormerod
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