Method for separating rare cells from a population of cells

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/04 (2006.01) C12Q 1/24 (2006.01) G01N 33/80 (2006.01) C12N 5/08 (2006.01)

Patent

CA 2182371

A method for isolating and enriching rare cells, including fetal nucleated erythrocytes from a peripheral blood sample is described. The method includes two centrifugation steps, the first a bulk separation step to enrich erythrocytes from other blood components. The second centrifugation comprises a colloidal density gradient medium dispersed in a hypertonic meltable gel. In one aspect of the invention, maternal erythrocytes may be hemolyzed prior to the second density gradient centrifugation to provide additional enrichment for fetal nucleated erythrocytes. The figure shows a series of graphs showing mean cell volume versus fraction number with and without chlorpromazine.

Procédé d'isolation et d'enrichissement de cellules rares, y compris d'érythrocytes nucléés foetaux, à partir d'un échantillon de sang périphérique. Ledit procédé comporte deux phases de centrifugation, la première étant une phase de séparation en masse destinée à enrichir les érythrocytes en les séparant d'autres composants sanguins. La seconde phase comporte un milieu colloïdal de centrifugation en gradient de densité dispersé dans un gel fusible hypertonique. Selon un aspect de la présente invention, des érythrocytes maternels peuvent être hémolysés avant la seconde centrifugation en gradient de densité pour obtenir un enrichissement supplémentaire des érythrocytes nucléés foetaux. La figure présente une série de graphiques montrant le volume cellulaire moyen par rapport au nombre fractionnaire avec et sans chlorpromazine.

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