Pellets used in cell culture and methods of making thereof

Abstract

The present invention is directed generally to dry cell culture media or feeds in pellet formats which can be reconstituted into liquid media for culturing cells in vitro. Each pellet composition may comprise the same or a different composition; for example, different vitamins, amino acids, buffers, trace salts, pH, iron chelators, etc. The invention also relates to methods of making dry cell culture media by altering ratios of different pellet compositions, or, methods of making modular dry cell culture media, or customizing media formulations for growing a cell type using pellets. The media pellets may be easier to handle either before reconstitution, during shipping and handling; and/or during reconstitution. Media pellets may be used in any container like bags including sterile, single use bags for preparing media formulations. The invention also relates to kits and culture systems using media pellets.

1 .- 26 . (canceled) 
     
     
         27 . A pelletized cell culture medium, feed, supplement or additive comprising one or more amino acids, one or more binders, one or more vitamins, one or more salts, and one or more trace components, wherein the amino acids are 25 to 40 percent, the binders are 20 to 65 percent, the vitamins are 1 to 5 percent, the salts are 2 to 10 percent, and the trace components are 0.01 to 0.05 percent. 
     
     
         28 . (canceled) 
     
     
         29 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the vitamins are selected from one or more of vitamin B12, biotin, choline, folic acid, niacinamide, pyridoxine, riboflavin, thiamine, ascorbic acid, and para-aminobenzoic acid (PABA). 
     
     
         30 . (canceled) 
     
     
         31 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the salts are selected from one or more of buffer salts, iron, zinc, calcium, copper, magnesium, manganese, ammonium, and vanadium salts. 
     
     
         32 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the amino acids are selected from one or more of the twenty amino acids, their salts or derivatives thereof. 
     
     
         33 . The pelletized cell culture medium, feed, supplement or additive of  claim 32 , wherein the amino acids are selected from one or more of glycine, alanine, arginine, aspartic acid, glutamic acid, histidine, isoleucine, methionine, phenylalanine, proline, hydroxyproline, serine, threonine, tryptophan, valine, tyrosine, cysteine and lysine. 
     
     
         34 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the binders are selected from the group consisting of a sugar, a natural substance, a synthetic substance, a semisynthetic substance, a microcrystalline cellulose, a glucose, a sucrose, a trehelose, a monosaccharide, a disaccharide, an oligosaccharide, an excipients and a disintegrant. 
     
     
         35 . The pelletized cell culture medium, feed, supplement or additive of  claim 34 , wherein the binders are microcrystalline cellulose and glucose. 
     
     
         36 . (canceled) 
     
     
         37 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the percent composition of binders in the pellet is from about 30 to about 60%. 
     
     
         38 . (canceled) 
     
     
         39 . The pelletized cell culture medium, feed, supplement or additive of  claim 27 , wherein the amino acid are 31-32 percent, the binder are 59-60 percent, and the salt are 6-7 percent. 
     
     
         40 . The pelletized cell culture medium, feed, supplement or additive of  claim 39  wherein the binder is D-glucose. 
     
     
         41 .- 58 . (canceled) 
     
     
         59 . A kit comprising:
 i) a first container comprising one or more of a cell culture medium, feed, supplement or additive of  claim 27 ,  29 ,  31  to  35 ,  37 ,  39  or  40 ;   ii) instructions for using the cell culture medium, feed, supplement or additive, wherein the cell culture medium, feed, supplement or additive is a pellet.   
     
     
         60 . A system comprising a liquid medium reconstituted from the pellet of  claim 59 . 
     
     
         61 . The system of  claim 60  wherein the cell in the cell culture is selected from the group consisting of a eukaryotic cell, a prokaryotic cell, an animal cell, a plant cell, a fungal cell, an algal cell, an insect cell, and a yeast cell, or wherein the cell is used for the cultivation of a virus or a viral particle. 
     
     
         62 . The system of  claim 60  or  61 , wherein the reconstituted liquid medium is used to cultivate a cell that produces a recombinant polypeptide, a virus, a secreted protein, or a cell either in suspension or an attached cell.