Abstract The viability of leukocytes or specific fractions of leukocytes is maintained by drawing blood from a donor or patient into a mixture of an anticlotting agent and a basic salt solution (BSS) or a minimum essential medium (MEM) which solution or medium sustains the viability of the leukocytes in the blood, then layering the blood-MEM or BSS mixture on a Ficoll-Hypaque solution and centrifuging to separate the leukocytes. The resulting leukocytes are diluted to about 0.5×10 6 viable cells per ml. in BSS or MEM containing antibiotics and fetal calf serum or serum substitutes equivalent thereto. The resulting suspension is transferred into sterile containers at room temperature so that each container has about 2-10×10 6 cells. The containers are then packed into insulated containers and are ready for shipping.
A method of maintaining the viability of leukocytes which comprises (a) drawing blood form a patient or donor directly into a mixture of an anticlotting agent and a medium which sustains the viability of leukocytes, (b) separating the leukocytes by centrifugation onto a solution having a specific gravity of about the same as the leukocytes being separated containing a mixture of a water soluble synthetic copolymer of sucrose and epichlorohydrin with an average molecular weight of 400,000±100,000 and sodium ditrizoate, (c) collecting the leukocytes and placing them in containers having a medium which sustains the viability of the leukocytes, said medium containing antibiotics and a substance selected from the group consisting of heat-inactivated fetal calf serum and a serum substitute equivalent thereto.
Claim 2
Independent Claim
122 words
A method of maintaining the viability of leukocytes which comprises (a) drawing blood form a patient or donor directly into a mixture of an anticlotting agent and a medium which sustains the viability of leukocytes, (b) separating the leukocytes by centrifugation onto a solution having a specific gravity of about the same as the leukocytes being separated containing a mixture of a water soluble synthetic copolymer of sucrose and epichlorohydrin with an average molecular weight of 400,000±100,000 and sodium ditrizoate, (c) collecting the leukocytes and placing them in containers having a medium which sustains the viability of the leukocytes, said medium containing antibiotics and a substance selected from the group consisting of heat-inactivated fetal calf serum and a serum substitute equivalent thereto.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein the anticlotting agent is heparin.
Claim 4
Dependent Claim
19 words
The method of claim 1 wherein the medium which sustains the viability of leukocytes is a basic salt solution.
Claim 5
Dependent Claim
19 words
The method of claim 1 wherein the medium which sustains the viability of leukocytes is a minimum essential medium.
Claim 6
Dependent Claim
25 words
The method of claim 1 wherein the leukocytes are lymphocytes and the specific gravity of the solution in step (b) is about 1.07 to 1.08.