Media and methods for enhancing the survival and proliferation of stem cells

Abstract

The present disclosure relates to improved supplements, culture media and methods for enhancing the survival or proliferation of mammalian stem cells. In particular, adding a lipid supplement, such as a lipid-enriched carrier (e.g. a lipid-enriched albumin), to the culture medium may enhance the survival and/or proliferation of the stem cells by at least 5% to 65% as compared to a culture medium that does not contain the lipid supplement.

1 . A culture medium for enhancing the survival or proliferation of one or more mammalian stem cells, comprising one or more lipids and a carrier,
 wherein the one or more lipids is one or both of oleic acid and palmitic acid, and   wherein an amount of oleic acid and/or palmitic acid is greater than an amount of alpha-linolenic acid and/or arachidonic acid.   
     
     
         2 . The culture medium according to  claim 1 , wherein the one or more lipids is selected from the group consisting of Mead's acid, arachidic acid, palmitoleic acid, myristic acid, myristoleic acid, linoleic acid, stearic acid, cholesterol, DL-alpha-tocopheryl and Kolliphor P188. 
     
     
         3 . The culture medium according to  claim 1 , wherein the carrier is an albumin, a liposome, an extracellular vesicle, an exosome, a nanostructured lipid carrier, or a cyclodextrin. 
     
     
         4 . The culture medium according to  claim 1 , wherein a concentration of the one or more lipids ranges from 1 ng/mL to 35 μg/mL. 
     
     
         5 . The culture medium according to  claim 1 , further comprising one or more survival factors. 
     
     
         6 . The culture medium according to  claim 5 , wherein the one or more survival factors comprise one or more small molecule inhibitors. 
     
     
         7 . The culture medium according to  claim 6 , wherein the one or more small molecule inhibitors comprise a Rho/Rock pathway inhibitor. 
     
     
         8 . The culture medium according to  claim 7 , wherein the Rho/Rock pathway inhibitor is Y-27632 at a concentration ranging between 1 μM to 20 μM. 
     
     
         9 . The culture medium according to  claim 1 , further comprising an extracellular matrix at a concentration below a gelation threshold thereof. 
     
     
         10 . The culture medium according to  claim 9 , wherein the gelation threshold is about 0.5% or higher. 
     
     
         11 . The culture medium according to  claim 10 , wherein the extracellular matrix comprises one or more monomatrix components, and the one or more matrix proteins comprise a: fibronectin, collagen, laminin, elastin, vitronectin, entactin, heparin sulphate, or proteoglycan. 
     
     
         12 . A method for enhancing the survival or proliferation of one or more mammalian stem cells, the method comprising culturing the one or more mammalian stem cells in a culture medium as claimed in  claim 1 . 
     
     
         13 . The method according to  claim 12 , wherein culturing comprises culturing as a monolayer. 
     
     
         14 . The method according to  claim 12 , wherein culturing comprises culturing the one or more cells under non-adherent conditions. 
     
     
         15 . The method according to  claim 12 , further comprising seeding the one or more cells at a seeding density of 1 cell/well up to about 1000 cells/cm 2 . 
     
     
         16 . The method according to  claim 12 , further comprising yielding a survival rate of about 5% to about 65% for the one or more cells. 
     
     
         17 . The method according to  claim 12 , further comprising exposing the one or more mammalian stem cells to an extracellular matrix. 
     
     
         18 . The culture medium according to  claim 17 , wherein a concentration of the extracellular matrix in the medium is below a gelation threshold thereof. 
     
     
         19 . The method according to  claim 18 , wherein the gelation threshold is about 0.5% or higher. 
     
     
         20 . The method according to  claim 19 , wherein the extracellular matrix comprises one or more monomatrix components, and the one or more matrix proteins comprise a: fibronectin, collagen, laminin, elastin, vitronectin, entactin, heparin sulphate, or proteoglycan.