Separation of viable from radiation-induced apoptotic lymphocytes by free-flow electrophoresis.
A human lymphocyte population undergoing apoptosis in vitro due to gamma-irradiation was fractionated by free-flow electrophoresis in triethanolamine--Na-acetate buffers, containing up to 50 mM NaCl, with pH 6.0, 7.2 and 8.5, made isotonic by addition of sucrose. As shown by a flow cytometric analysis of the eluate, the distribution of apoptotic lymphocytes is shifted to the range of higher electrophoretic mobilities relative to that of viable ones at pH 8.5, yielding cell fractions enriched in apoptotic cells by a factor of 3 to 5. The difference in rates of electrophoretic migration observed at a mildly alkaline pH but not at a neutral or mildly acidic one suggests that the surface of apoptotic lymphocytes is more acidic than that of viable ones.
Chestkov
V
V
Section on Macromolecular Analysis, Laboratory of Cellular and Molecular Biophysics, NICHD, National Institutes of Health, Bethesda, MD 20892-1580, USA.
Baibakov
B
B
Radko
S P
SP
Chrambach
A
A
D017209:Apoptosis / Q000528:radiation effects; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival / Q000528:radiation effects; D004586:Electrophoresis / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008214:Lymphocytes / Q000528:radiation effects