Cellular microarrays for screening differentiation factors

Abstract

Provided is a microarray platform for the culture of cells atop combinatorial matrix mixtures; enabling the study of differentiation in response to a multitude of microenvironments in parallel.

1 . A cell culture substrate, comprising a plurality of microspot islands or microwells in an array, wherein the microspot islands or microwells comprise an insoluble factor or an insoluble and soluble factor, wherein the insoluble factor promotes cellular adhesion. 
     
     
         2 . The cell culture substrate of  claim 1 , wherein the substrate comprises a polymerized biopolymer. 
     
     
         3 . The cell culture substrate of  claim 2 , wherein the biopolymer is a hydrogel or a relatively homogeneous slab of biopolymer. 
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . The cell culture substrate of  claim 1 , further comprising a plurality of microfluidic channels connecting one or more microspot islands or microwells to a fluid flow. 
     
     
         7 . A method of making a microarray, comprising:
 spotting a plurality of locations on a substrate with an adherence material.   
     
     
         8 . The method of  claim 7 , wherein the adherence material is an insoluble adherence material or an extracellular matrix protein. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 7 , wherein the substrate is layered with a hydrogel and wherein the hydrogel is etched at each of the plurality of locations to form a microwell. 
     
     
         11 . (canceled) 
     
     
         12 . The method of  claim 7 , wherein the substrate further comprises fluid flow channels fluidly connecting each location with a fluid flow. 
     
     
         13 . The method of  claim 7 , further comprising providing one or more cell types that adhere to each location comprising the adherence material. 
     
     
         14 - 17 . (canceled) 
     
     
         18 . The method of  claim 10 , wherein the hydrogel is a polyacrylamide gel. 
     
     
         19 . The method of  claim 7 , wherein the spotting is performed by a spotting device. 
     
     
         20 . A method of making a culture substrate, comprising:
 spotting a material on the substrate using a device capable of spotting from about 1 to about 1000 nanoliters of material to generate an island of material.   
     
     
         21 . The method of  claim 20 , wherein the device is a DNA spotting device. 
     
     
         22 . The method of  claim 20 , wherein the material comprises an insoluble factor or a printing buffer. 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 7 , wherein the printing buffer comprises 100 mM acetate, 4 mM EDTA, 20% glycerol and 0.25% Triton X-100 at pH 5.0. 
     
     
         25 - 28 . (canceled) 
     
     
         29 . A culture system comprising the microarray of  claim 7  and one or more cell-types. 
     
     
         30 . An assay system comprising:
 contacting a microarray of  claim 7  with one or more cell-types and measuring an activity selected from gene expression, cell function, metabolic activity, morphology, and a combination thereof.   
     
     
         31 . The assay system of  claim 30 , wherein the system further comprising contacting the cells with a test agent once the cells are in stable culture on the substrate. 
     
     
         32 . The assay system of  claim 30 , wherein a factor is present in a material of an island on the substrate. 
     
     
         33 . The method of  claim 7 , further comprising providing one or more soluble or insoluble material/biological material that adhere to each location comprising the adherence material.