Media for Cell Culture

Abstract

The present disclosure relates, in general, to a media, e.g., a serum replacement, media supplement, complete media or cryopreservation media, comprising a base physiological buffer and liposomes comprising cholesterol, phosphatidylcholine and fatty acids. It is contemplated that media provides advantages to improve cell growth in culture compared to cells cultured not using the serum replacement described herein.

1 . A medium for use with cells in suspension or in adherent culture, the media comprising a base physiological buffer liquid mix and
 (a) liposomes comprising cholesterol, phosphatidylcholine and fatty acids, wherein the liposome is in an amount such that the final concentration of cholesterol in a cell suspension or adherent culture is from 1 to 20 mg/L, and wherein the final concentration of phosphatidylcholine in a cell suspension or adherent culture is from 100 to 1000 mg/L; and   (b) pectin.   
     
     
         2 . The medium of  claim 1 , wherein the liposome comprises one or more fatty acids selected from the group consisting of linolenic acid, linoleic acid, myristic acid and oleic acid. 
     
     
         3 . (canceled) 
     
     
         4 . The medium of  claim 1 , wherein the final concentration of pectin in a cell suspension or adherent culture is from 25 to 500 mg/L. 
     
     
         5 . The medium of  claim 1 , wherein the base physiological buffer liquid mix comprises one or more organic salt, inorganic salt, buffer, iron source or iron transporter, glycerol, amino acid, vitamin, sugar, antioxidant and trace element. 
     
     
         6 - 10 . (canceled) 
     
     
         11 . The medium of  claim 5 , wherein the medium comprises one or more trace element selected from the group consisting of selenium, molybdnate, chromium, cobalt, nickel, zinc, copper, manganese, barium, gallium, lithium, tin, titanium, bromium, iodine, vanadium, germanium, molybdenum, silicon, iron, fluorine, silver, rubidium, zirconium, cadmium, and aluminum. 
     
     
         12 . The medium of  claim 11 , wherein the trace element is selenium and the final concentration of selenium in a cell suspension or adherent culture is from 0.005 mg/L to 0.05 mg/L. 
     
     
         13 . The media of  claim 1 , wherein the media is a serum replacement, complete media, media supplement or cryopreservation media. 
     
     
         14 . A serum replacement, complete media or media supplement comprising liposomes and a base physiological buffer liquid mix, wherein the liposomes comprise phosphatidylcholine, ethanolamine, linolenic acid, linoleic acid, cholesterol, and polysorbate, and the base liquid mix comprises at least one inorganic salt, at least one sugar, glycerol and at least one trace element. 
     
     
         15 . The serum replacement, complete media or media supplement of  claim 14  further comprising at least one non-ionic surfactant and at least one iron source. 
     
     
         16 . The serum replacement, complete media or media supplement of  claim 14 , wherein the liposome is in an amount such that the final concentration of cholesterol in a cell suspension or adherent culture is from 1 to 20 mg/L, and wherein the final concentration of phosphatidylcholine in a cell suspension or adherent culture is from 100-1000 mg/L. 
     
     
         17 - 18 . (canceled) 
     
     
         19 . The serum replacement, complete media or media supplement of  claim 14 , wherein the liquid mix comprises one or more trace elements selected from the group consisting of selenium, molybdnate, chromium, cobalt, nickel, zinc, copper, manganese, barium, gallium, lithium, tin, and titanium. 
     
     
         20 - 22 . (Canceled) 
     
     
         23 . The serum replacement, complete media or media supplement of  claim 14 , wherein the liquid mix comprises one or more non-ionic surfactants selected from the group consisting of Pluronic-68, F68 Pastille, Pluronic-128, sorbitan, polysorbate, and block copolymers. 
     
     
         24 . (canceled) 
     
     
         25 . A serum replacement of  claim 14 , wherein the final concentration of cholesterol in the serum replacement is from about 10 to 200 mg/L and the final concentration of the phosphotidylcholine in the serum replacement is from about 1000 mg to 10 g/L. 
     
     
         26 . A complete media of  claim 14 , wherein the concentration of cholesterol in the complete media is from about 1 to 20 mg/L and the concentration of the phosphotidylcholine in the complete media is from about 100 mg to 1000 mg/L. 
     
     
         27 . A media supplement of  claim 14 , wherein the concentration of cholesterol in the media supplement is from about 100 to 2000 mg/L and the concentration of the phosphotidylcholine in the media supplement is from about 10 g to 100 g/L. 
     
     
         28 . The serum replacement, complete media or media supplement of  claim 14 , further comprising pectin. 
     
     
         29 . The media, serum replacement, complete media or media supplement of  claim 14 , wherein the liposome is a nanoparticle. 
     
     
         30 - 31 . (canceled) 
     
     
         32 . A cell cryopreservation media comprising liposomes and a base physiological buffer liquid mix, wherein the final concentration of cholesterol in the cryopreservation media is from about 10 to 200 mg/L and the final concentration of the phosphotidylcholine in the cryopreservation media is from about 1000 mg to 10 g/L. 
     
     
         33 . The cryopreservation media of  claim 32  further comprising pectin. 
     
     
         34 . (canceled) 
     
     
         35 . The cryopreservation media of  claim 32 , further comprising glycerol. 
     
     
         36 . (canceled) 
     
     
         37 . The cryopreservation media of  claim 32 , further comprising dimethyl sulfoxide (DMSO), wherein the DMSO is at a concentration of less than 4%. 
     
     
         38 . The cryopreservation media of  claim 32  wherein the media further comprises polylysine. 
     
     
         39 . A method of culturing cells comprising culturing cells in a media containing a serum replacement, complete media or media supplement of  claim 14 . 
     
     
         40 - 41 . (canceled) 
     
     
         42 . A method of cryopreserving cells comprising suspending cells in a cryopreservation media of  claim 32  and placing the cells in an environment of less than 8° C. 
     
     
         43 - 46 . (canceled)