Human plasma-like medium

Abstract

In some aspects, described herein are cell culture media that are useful for in vitro culture of mammalian cells. The culture media contain a variety of small organic compounds that are found in normal adult human blood. Also described are methods of using the culture media for a variety of purposes. Also described are methods of treating cancer.

1 . A basal culture medium, comprising:
 at least 9 proteinogenic amino acids;   one or more vitamins;   one or more inorganic ions;   glucose; and   at least 10 small organic compounds selected from 4-hydroxyproline, acetylglycine, alpha-aminobutyrate, betaine, carnitine, citrulline, ornithine, taurine, 2-hydroxybutyrate, 3-hydroxybutyrate, acetate, citrate, formate, lactate, malonate, pyruvate, succinate, acetone, creatine, creatinine, glutathione, glycerol, urea, galactose, fructose, hypoxanthine, and uric acid.   
     
     
         2 - 43 . (canceled) 
     
     
         44 . A method of culturing one or more mammalian cells comprising providing the basal culture medium of  claim 1  or the culture medium of  claim 1  further comprising serum or a serum substitute; and culturing one or more mammalian cells in the culture medium. 
     
     
         45 . (canceled) 
     
     
         46 . The method of  claim 44 , wherein the one or more mammalian cells comprise blood cells. 
     
     
         47 . The method of  claim 44 , wherein the one or more mammalian cells comprise cancer cells. 
     
     
         48 . The method of  claim 44 , further comprising adding a test agent to the culture medium. 
     
     
         49 . A method of characterizing a mammalian cell population or cell line comprising: (a) culturing one or more cells of a mammalian cell population or cell line in the the basal culture medium of  claim 1  or the culture medium of  claim 1  further comprising serum or a serum substitute; and (b) detecting a phenotype of the cells cultured in the culture medium or obtaining results of an assay performed with the cells cultured in the culture medium. 
     
     
         50 . The method of  claim 49 , further comprising (c) culturing one or more cells from the same population or cell line in a second culture medium; (d) detecting a phenotype of the cells cultured in the second culture medium or obtaining results of an assay performed with the cells cultured in the second culture medium; and (e) comparing the phenotype detected in step (b) with the phenotype of one or more cells from the same population or cell line that have been cultured in a second culture medium or comparing results of the assay with results of the same assay performed with one or more cells from the same population or cell line that have been cultured in a second culture medium. 
     
     
         51 . The method of  claim 49 , wherein the method comprises detecting a difference between the phenotype or assay results obtained from cells cultured in the first culture medium and the phenotype or assay results obtained from cells cultured in the second culture medium. 
     
     
         52 . The method of  claim 49 , wherein the method comprises contacting the cells with a test agent during part or all of the period during which they are cultured in the culture medium. 
     
     
         53 . The method of  claim 52 , wherein the method comprises comparing the phenotype of cells cultured in the culture medium in the presence of the test agent with the phenotype of cells from the same population or cell line that have been cultured in the second culture medium in the presence of the test agent or comparing results of the assay performed with cells cultured in the culture medium in the presence of the test agent with results of the same assay performed with cells from the same population or cell line that have been cultured in the second culture medium in the presence of the test agent. 
     
     
         54 . (canceled) 
     
     
         55 . The method of  claim 49 , wherein the assay is a viability assay, proliferation assay, apoptosis assay, autophagy assay, reporter assay, or cytotoxicity assay. 
     
     
         56 . A method of characterizing a test agent comprising: (a) culturing one or more mammalian cells in the basal culture medium of  claim 1 , or the culture medium of  claim 1  further comprising serum or a serum substitute in the presence of the test agent; and (b) obtaining results of an assay performed with the cells. 
     
     
         57 . The method of  claim 56 , further comprising (c) culturing one or more cells from the same population or cell line in a second culture medium in the presence of the test agent and (d) obtaining results of an assay performed with the cells cultured in the second culture medium. 
     
     
         58 . The method of  claim 56 , further comprising comparing results of the assay with results of the same assay performed with one or more cells from the same population or cell line that have been cultured in a second culture medium in the presence of the test agent. 
     
     
         59 - 60 . (canceled) 
     
     
         61 . The method of  claim 56 , wherein the method comprises detecting a difference between the assay results obtained from cells cultured in the first culture medium in the presence of the test agent and the assay results obtained from cells cultured in the second culture medium in the presence of the test agent. 
     
     
         62 . (canceled) 
     
     
         63 . The method of  claim 49 , wherein the one or more cells comprise blood cells. 
     
     
         64 . The method of  claim 49 , wherein the one or more cells comprise cancer cells. 
     
     
         65 . The method of  claim 56 , wherein the assay is a viability assay, proliferation assay, apoptosis assay, autophagy assay, reporter assay, or cytotoxicity assay. 
     
     
         66 . The method of  claim 56 , wherein the test agent is a small molecule. 
     
     
         67 . The method of  claim 56 , wherein the test agent is a chemotherapeutic agent. 
     
     
         68 - 73 . (canceled)