Expansion medium for CD34-negative stem cells

A cell growth medium supplement comprising (a) human platelet lysate free of solid matter greater than 0.22 μm in diameter, wherein the lysate constitutes from 17% to 94% of the total volume of the cell growth medium supplement;

• and (b) human fresh frozen plasma (FFP) filtrate free of solid matter greater than 0.22 μm in diameter, wherein the FFP filtrate constitutes from 6% to 83% of the total volume of the cell growth medium supplement;
• wherein when the cell growth medium supplement is combined with heparin and an L-glutamine-containing, serum-free, low glucose medium suitable for mammalian cell growth, in order to form a cell growth medium containing (i) 3% to 25% by volume of the cell growth medium supplement, (ii) heparin at a concentration of from 0 U/ml to 10 U/ml, and (iii) L-glutamine at a concentration of from 0.5 mM to 10 mM, (i) wherein the FFP is the fluid portion of human blood that has been centrifuged, separated, and frozen solid within six hours of collection, (ii) wherein the resulting cell growth medium permits the expansion of human CD34−stem cells, and (iii) wherein the resulting expanded CD34−stem cells retain the ability to differentiate.

A kit for use in expanding human CD34− stem cells comprising, in separate compartments, (a) a human platelet lysate free of solid matter greater than 0.22 μm in diameter, and (b) a human fresh frozen plasma (FFP) filtrate free of solid matter greater than 0.22 μm in diameter, wherein (i) the lysate constitutes from 17% to 94% of the combined volume of (a) and (b), (ii) the FFP filtrate constitutes from 6% to 83% of the combined volume of (a) and (b), and (iii) when the lysate and filtrate are combined with heparin and with L-glutamine-containing, serum-free, low glucose medium suitable for mammalian cell growth, in order to form a cell growth medium wherein (i) the kit's contents constitute 3% to 25% of the medium by volume and (ii) heparin is at a concentration of from 0 U/ml to 10 U/ml, (i) wherein the FFP is the fluid portion of human blood that has been centrifuged, separated, and frozen solid within six hours of collection, (ii) wherein the resulting cell growth medium permits the expansion of human CD34−stem cells, and (iii) the resulting expanded CD34−stem cells retain the ability to differentiate.