Functionalized substrate to manipulate cell function and differentiation

Abstract

The invention relates to a scaffold for steering cells into a predetermined direction of cell functionality, preferably a cell differentiation scaffold for steering cells into a predetermined direction of cell differentiation. The scaffold comprises a polydimethylsiloxane (PDMS)-, or rubber-, or silicone-based polymeric surface, and one or more cell functionality-inducing stimuli, preferably one or more cell differentiation-inducing stimuli, coupled to the polymeric surface.

1 . A substrate, for steering of cells into a predetermined direction of cell functionality, the steering of cells consisting of steering cells into a predetermined direction of cell differentiation, the substrate comprising:
 an elastomer material having coupled thereto one or more cell functionality-inducing stimuli wherein the cell functionality-inducing stimuli comprise cell differentiation-inducing stimuli.   
     
     
         2 . The substrate according to  claim 1 , wherein the one or more cell functionality-inducing stimuli are selected from the group consisting of: cell receptor ligands, ectracellular matrix modules, artificial receptor agonists or antagonists, and specific antigens for T or B cell receptors. 
     
     
         3 . The substrate according to  claim 1 , wherein the one or more cell functionality-inducing stimuli are cell receptor ligands. 
     
     
         4 . The substrate according to  claim 1 , wherein the cell receptor ligands are configured for activating signaling through a signaling pathway selected from the group consisting of: nuclear receptors, such as ER, AR; progesterone receptors; growth factor pathways, such as: PI3K-FOXO, JAK-STAT3, MAPK-AP1 pathways; immune pathways, such as STAT1/2 type I interferon (STAT1/2-1), STAT1/2 type II interferon (STAT1/2-2) pathways; developmental pathways, such as Hedgehog, Notch, TGF-β, and Wnt signaling pathways; and inflammatory pathways such as NFkB. 
     
     
         5 . The substrate according to  claim 1  wherein the elastomer material comprises one or more polysiloxane polymers or polybutadiene polymers. 
     
     
         6 . The substrate according to  claim 1 , wherein the one or more cell functionality induced stimuli coupled to the elastomer material are obtainable by reacting a functional group of the one or more cell functionality-induced stimuli with a further reactive group of a precursor elastomer material using a crosslinking reaction chosen from the group of bioconjugation reactions. 
     
     
         7 . The substrate according to  claim 1 , wherein the one or more cell functionality-inducing stimuli are coupled to the elastomer material via one or more of an amide, amine and ester group. 
     
     
         8 . A culture unit comprising a cell culture chamber having therein a substrate according to  claim 1 . 
     
     
         9 . Use of the substrate or culture unit according to  claim 8 , for producing a cell culture tissue or organ. 
     
     
         10 . An In vitro method of producing a cell culture, tissue or organ, the method comprising:
 (a) determining first culture conditions that control activity of at least one first signaling pathway of cells to be cultivated, the activity of the at least one first signaling pathway being a selected activity to steer the cells into a first predetermined direction of cell functionality; and   (b) cultivating cells in a culture unit under the first culture conditions to obtain a cell culture.   
     
     
         11 . The In vitro method according to  claim 10 , further comprising:
 (c) inferring activity of the at least one first signaling pathway in a cell sample obtained from the cell culture;   (d) comparing the inferred activity with the selected activity, and   (e) determining second culture conditions based on the comparison of the inferred activity with the selected activity.   
     
     
         12 . The In vitro method according to  claim 11 , further comprising determining second culture conditions based on the comparison of the inferred activity with the selected activity and wherein the second culture conditions are determined to (i) reduce deviation of the inferred activity from the selected activity, or (ii) control a selected activity of at least a second signaling pathway different from the at least one first signaling pathway, the selected activity of the at least one second signaling pathway being selected to steer the cells into a second predetermined direction of cell functionality different from the first predetermined direction of cell functionality. 
     
     
         13 . The In vitro method according to  claim 10 , further comprising:
 (f) cultivating the cells in the culture unit under the second culture conditions;   (g) inferring activity of the at least one first signaling pathway in a cell sample obtained from the cell culture;   (h) comparing the inferred activity with the selected activity, and   (i) determining third culture conditions based on the comparison of the activity inferred in steps (g) and (h) with the selected activity.   
     
     
         14 . The In vitro method according to  claim 9 ,
 wherein the cell functionality is selected from the group consisting of cell differentiation, maintenance of pluripotency and cellular function, such as cell division, migration and metabolism, , adhesion, apoptosis, generating oxidative stress, DNA repair, production of secretory proteins, production of extracellular matrix proteins, preferably cell differentiation.   
     
     
         15 . The In vitro method according to  claim 9 ,
 wherein the first and/or second culture conditions are defined by controlled exposure of the cells to one or more cell functionality-inducing stimuli selected from the group consisting of cell receptor ligands, extracellular matrix molecules, artificial receptor agonists or antagonists, and specific antigens for T or B cell receptors, the stimuli controlling activity of the at least one first and/or second signaling pathway, and/or   wherein the one or more cell functionality-inducing stimuli are provided in the form of one or more culture units.   
     
     
         16 . The In vitro method according to  claim 9 ,
 wherein the cells are mouse stem cells, human stem cells, pluripotent stem cells, mesenchymal stem cells, embryonic stem cells, adult stem cells, tumor cells, cell lines.   
     
     
         17 . A non-transitory storage medium storing instruction that are executable by a digital processing device to perform the method of  claim 9 . 
     
     
         18 . A computer program comprising program code means for causing a digital processing device to perform the method of  claim 9 , when the computer program is run on a digital processing device. 
     
     
         19 . A system for performing the method of  claim 14 , the system comprising
 (a) one or more culture units,   (b) a sensor unit for inferring an activity of at least one first signaling pathway;   (c) a controller that is configured to compare the inferred activity with a predetermined activity of the at least one first signaling pathway, the predetermined activity being selected so to controllably steer the cells into a first predetermined direction of cell functionality, and   (d) a computer program,
 wherein, preferably, the controller is further configured to determine culture conditions based on the comparison of the inferred activity with the predetermined activity, the culture conditions being optionally determined so as to (ii) reduce deviation of the inferred activity from the predetermined activity or (ii) control activity of at least one second signaling pathway, the activity of the at least one second signaling pathway.