A process for obtaining intact and viable leucocytes and thrombocytes from blood

Claim 2

A process for recovering leucocytes and thrombocytes from blood, which comprises: a) contacting the blood with an erythrocyte-aggregating substance to produce a liquid component and aggregated erythrocytes and separating the aggregated erythrocytes from the liquid component; b) centrifuging the liquid component at a speed having an average force of at most about 400 x g and at a temperature of about 0 to 37°C for about 5 to 15 min to produce a thrombocyte-containing supernatant and sedimented leucocytes and separating the sedimented leucocytes from the supernatant; c) suspending the sedimented leucocytes in water or up to a 0.4% sodium chloride solution at a pH of about 6.0 to 8.5, and at a maximum temperature of about +8°C for about 20 to 60 seconds, and thereafter readjusting the suspension to physiological concentration and separating the leucocytes from the suspension.

Claim 3

A process according to claim 1, which further comprises suspending the leucocytes from step c at least once in a liquid medium that does not damage or aggregate leucocytes and then separating the leucocytes from the liquid medium.

Claim 4

A process according to claim I wherein methylcellulose of low viscosity is used as the erythrocyte-aggregating substance.

Claim 5

A process according to claim 3 wherein methylcellulose with a viscosity of (2-3)x 10 -2 Pa.s measured in a 2% solution in water at 20°C is used.

Claim 6

A process according to claim 3 wherein the methylcellulose is used in a concentration of 0.05% to 0.3%.

Claim 7

A process according to claim 1 wherein the centrifugation of step b is carried out at an average force of 400 x g and a maximum temperature of 15°C for about 8 to 12 min.

Claim 8

A process according to claim 1 wherein step c is performed at a pH of about 6.8 to 7.5, a maximum temperature of 4°C, and an NaCl concentration of about 0.2% for about 25 to 35 sec.

Claim 9

A process according to claim 2 wherein a solution of physiological saline, preferably physiological sodium chloride, is used as the liquid medium that does not damage or aggregate leucocytes.

Claim 10

A process according to claim 1 which further comprises centrifuging the thrombocyte-containing supernatant of step b at a speed corresponding to an average force higher than 400 x g over a period of 20 min to produce sedimented thrombocytes and a second supernatant containing the plasma constituents.

Claim 11

A process according to claim 9 which further comprises suspending the sedimented thrombocytes at least once in a liquid medium that does not damage or aggregate thrombocytes and then separating the thrombocytes from the liquid medium.

Claim 12

A process according to claim I, which comprises: a) contacting the blood with methylcellulose at about 0° to about 37°C to produce a liquid component and aggregated erythrocytes and separating the aggregated erythrocytes from the liquid component, the methylcellulose having a viscosity of 2 to 3 x 10 -2 P a.s as measured in a 2% solution in water at 20°C and forming a concentration in the blood of 0.05% to 0.3%; b) centrifuging the liquid component at a speed having an average force of 400 x g and at a temperature of at most 15°C for about 8 to 12 minutes to produce a thrombocyte-containing supernatant and sedimented leucocytes and separating the sedimented leucocytes from the supernatant; c) suspending the sedimented leucocytes in a 0.2% sodium chloride aqueous solution at a pH of about 6.8 to 7.5 and at a temperature of at most 4°C for about 25 to 35 seconds, and thereafter readjusting the suspension to physiological concentration and separating the leucocytes from the suspension; d) centrifuging the thrombocyte-containing supernatant of step b at a speed corresponding to an average force of about 800 x g over a period of 20 minutes to produce sedimented thrombocytes and a second supernatant and separating the sedimented thrombocytes from the second supernatant.