Provision of density specific blood cells for the...

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G01N 33/49 (2006.01) C12Q 1/02 (2006.01) C12Q 1/24 (2006.01) G01N 1/34 (2006.01) G01N 33/50 (2006.01) G01N 33/574 (2006.01)

Patent

CA 2001177

An improved method is provided for the isolation of lymphocytes for use in the SCM test for the detection of cancer and other diseases and conditions. The method of the present invention can isolate either or both of the SCM-responding lymphocyte fractions from the same blood sample: the F2 fraction, with a buoyant density of 1.0590 g/cm3 to 1.0670 g/cm3 measured at 20°C in a solution having an osmolality of about 0.315 Osm/kg, and the F4 fraction, with a buoyant density of 1.0690 g/cm3 to 1.0730 g/cm3. These lymphocyte fractions are isolated in visible bands and are substantially free of lymphocytes having other buoyant densities. This avoids cross-contamination of the lymphocyte fractions with each other as well as with SCM non- responding lymphocytes. Several gradients useful in this isolation method are described. This isolation method can use as starting material either a blood sample depleted of phagocytic cells or the total population of lymphocytes. The latter avoids the use of iron powder or carbonyl-iron powder in removing the phagocytic cells.

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