Methods and apparatus for conditioning cell populations for cell therapies

Abstract

A bioreactor system for conditioning of pluripotent cells or cell media is provided. In further aspects, conditioned pluripotent cells and methods for making such cells are provided.

We claim: 
     
         1 . A method of producing a conditioned composition comprising:
 (a) culturing a population of stem cells on a substrate to allow cell adhesion; and   (b) applying a controlled shear stress to the stem cells with a force sufficient to produce a conditioned composition.   
     
     
         2 . The method of  claim 1 , wherein the method is automated. 
     
     
         3 . The method of  claim 1 , wherein the substrate is a surface that supports the growth of the stem cells in a monolayer. 
     
     
         4 . The method of  claim 1 , wherein a growth medium is passed over the stem cells. 
     
     
         5 . The method of  claim 1 , wherein the shear stress is applied in a bioreactor system. 
     
     
         6 . The method of  claim 1 , wherein the shear stress is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 dynes per square centimeter. 
     
     
         7 . The method of  claim 1 , wherein the shear stress is at least 5, 10 or 15 dynes per square centimeter. 
     
     
         8 . The method of  claim 7 , wherein the shear stress is at least 5 dynes per square centimeter. 
     
     
         9 . The method of  claim 7 , wherein the shear stress is at least 10 or 15 dynes per square centimeter. 
     
     
         10 . The method of  claim 1 , wherein the stem cells are human cells. 
     
     
         11 . The method of  claim 1 , wherein the stem cells are induced pluripotent stem (iPS) cells. 
     
     
         12 . The method of  claim 1 , wherein the stem cells are mesenchymal stem cells (MSCs). 
     
     
         13 . The method of  claim 12 , further comprising isolating the MSCs from a tissue. 
     
     
         14 . The method of  claim 13 , wherein the tissue is bone marrow, cord blood, peripheral blood, fallopian tube, fetal liver, lung, dental pulp, placenta, adipose tissue, or amniotic fluid. 
     
     
         15 . The method of  claim 1 , wherein the conditioned stem cells have at least 6-fold higher expression of an anti-inflammatory gene compared to the starting populating of stem cells. 
     
     
         16 . The method of  claim 1 , wherein the stem cells are autologous. 
     
     
         17 . The method of  claim 3 , wherein the substrate is glass or plastic. 
     
     
         18 . The method of  claim 17 , wherein the substrate is coated with at least one substance selected from collagen IV, fibronectin, laminin, vitronectin hydrocarbon polymers, metals and/or oxides. 
     
     
         19 . The method of  claim 4 , wherein the growth medium comprises at least one factor selected from IL1B, TNF-α, IFNγ, PolyI:C, lipopolysaccharide (LPS), phorbol myristate acetate (PMA) and/or a prostaglandin. 
     
     
         20 . The method of  claim 19 , wherein the prostaglandin is 16,16′-dimethyl prostaglandin E2 (dmPGE2).