Cell culture medium

Abstract

The present invention relates to a cell culture medium for culturing human cells comprising an anti-coagulated total blood material wherein the hemoglobin level is from about 8 to about 16 g/dl. More particularly, the invention provides a cell culture medium in which cells present in the blood are disrupted and the insoluble remnants of the lysated cells are removed. Further, the invention provides a method for the preparation of a cell culture medium for culturing human cells, according to the invention.

1 . A cell culture medium for culturing human cells comprising an anti-coagulated total blood material wherein the hemoglobin level is from about 8 to about 16 g/dl. 
     
     
         2 . The cell culture medium according to  claim 1 , wherein the cells present in the blood are disrupted or lysed and the insoluble remnants of the disrupted or lysed cells are removed. 
     
     
         3 . The cell culture medium according to  claim 1 , wherein the blood is of mammalian origin. 
     
     
         4 . The cell culture medium according to  claim 3 , wherein the blood is of human origin. 
     
     
         5 . The cell culture medium according to  claim 1 , wherein the ion content of the medium is restored to match that of the extracellular fraction of total blood starting material. 
     
     
         6 . The cell culture medium according to  claim 1 , wherein the low molecular weight (<3000 Dalton) nutrient content and/or composition is essentially the same as that of a medium selected form the group consisting of Roswell Park Memorial Institute (RPMI) medium, Dulbecco's Modified Eagle's Medium (DMEM), and Isocove's Modified Dulbecco's Medium (IMDM). 
     
     
         7 . The cell culture medium according to  claim 1 , wherein the reducing capacity of the medium is restored by addition of reduced glutathione. 
     
     
         8 . (canceled) 
     
     
         9 . A method for the preparation of the cell culture medium for culturing human cells of  claim 1 , said method comprising
 (a) providing anti-coagulated mammalian blood as starting material;   (b) mechanically disrupting or lysing the cells present in the blood;   (c) removing the insoluble remnants of the disurupted or lysed cells.   
     
     
         10 . The method according to  claim 9 , wherein the mechanical disruption of the cells is achieved by freezing the total blood or by ultrasound, electric blade homogenizer or French press. 
     
     
         11 . The method according to  claim 9 , wherein removal of the insoluble remnants of the lysated cells is achieved by ultracentrifugation or mechanical filtration. 
     
     
         12 . The method according to  claim 9 , wherein the low molecular weight (<3000 Dalton) content and/or composition of the medium is adjusted by dialysis against conventional cell culture medium selected form the group consisting of RPMI, DMEM, and IMDM. 
     
     
         13 . The method according to  claim 9 , wherein the reducing capacity of the medium is restored by addition of reduced glutathione. 
     
     
         14 . Cell culture medium for culturing human cells, obtainable by the method according to  claim 9 . 
     
     
         15 . (canceled) 
     
     
         16 . The cell culture medium according to  claim 1 , wherein said culture medium is prepared by
 (a) obtaining mammalian blood having an extracellular fraction, as starting material;   (b) adding an anticoagulant to the mammalian blood thereby providing anti-coagulated mammalian blood;   (c) mechanically disrupting or lysing the cells present in the anti-coagulated mammalian blood;   (d) removing insoluble remnants of the disrupted or lysed cells to produce a particle-free extract;   (e) modifying the ion content of the particle-free extract to match the extracellular fraction of the starting material or to be similar to that of a medium selected form the group consisting of Dulbecco's Modified Eagle's Medium (DMEM), Roswell Park Memorial Institute (RPMI) medium and Isocove's Modified Dulbecco's Medium (IMDM).   
     
     
         17 . The cell culture medium according to  claim 16 , wherein the growth and survival factor flora of the starting material is preserved in the culture medium. 
     
     
         18 . The cell culture medium according to  claim 16 , wherein the anticoagulant is heparin. 
     
     
         19 . The cell culture medium according to  claim 16 , wherein the disrupting or lysing of the cells achieves an even disruption of cell membranes of the cells. 
     
     
         20 . The cell culture medium according to  claim 16 , wherein the culturing medium contains all macromolecular components of the blood. 
     
     
         21 . The cell culture medium according to  claim 16 , wherein the mechanical disruption or lysing of the cells is achieved by freezing the total blood.