Three-dimensional fibrous scaffolds for cell culture

Abstract

Provided herein is a three-dimensional scaffold composition comprising randomly oriented fibers, wherein the fibers comprise a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) and a poly(lactic-co-glycolic acid) (PLGA). Also provided are methods for using the three-dimensional scaffolds described herein.

What is claimed: 
     
         1 . A composition comprising:
 (i) a three-dimensional scaffold of randomly oriented fibers, wherein said fibers are electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) and a poly(lactic-co-glycolic acid) (PLGA); and   (ii) a population of cells isolated from a tumor of an animal or human, wherein said population of cells is inoculated onto said scaffold without prior incubation of the cells in a cell culture medium.   
     
     
         2 . The composition of  claim 1 , wherein the ratio of PEG to PLA is from about 1:2 to about 1:20. 
     
     
         3 . The composition of  claim 2 , wherein the ratio of PEG to PLA is from about 1:4 to about 1:10. 
     
     
         4 . The composition of  claim 3 , wherein fiber diameter ranges from about 0.3 μm to about 10 μm. 
     
     
         5 . The composition of  claim 1 , wherein the scaffold comprises pores having a diameter between about 5 mm to about 20 μm. 
     
     
         6 . The composition of  claim 5 , wherein the scaffold comprises pores having a diameter of less than about 10 μm. 
     
     
         7 . The composition of  claim 1 , wherein the PEG has a molecular weight of about 2 kDa. 
     
     
         8 . The composition of  claim 1 , wherein the PLGA has a lactic acid:glycolic acid ratio of between about 75:25 to about 95:. 
     
     
         9 . The composition of  claim 8 , wherein the PLGA has a lactic acid:glycolic acid ratio of about 85:15. 
     
     
         10 . The composition of  claim 1 , wherein the population of cells comprises a population of cancerous cells and stromal cells isolated from a tumor of an animal or human subject. 
     
     
         11 . The composition of  claim 1 , wherein said composition is incubated in a cell culture medium, and wherein the population of cells is a tumoroid. 
     
     
         12 . The composition of  claim 11 , wherein the tumoroid comprises a population of cells having increased resistance to a therapeutic agent compared to the population of cells isolated from the tumor of the animal or human subject but cultured in the absence of the scaffold. 
     
     
         13 . A composition incubated in a cell culture medium comprising:
 (i) a three-dimensional scaffold of randomly oriented fibers, wherein said fibers are electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) having a ratio of PEG to PLA of from about 1:4 to about 1:10, and a poly(lactic-co-glycolic acid) (PLGA) having a lactic acid:glycolic acid ratio of about 85:15, and wherein fiber diameter ranges from about 0.3 μm to about 10 μm, the scaffold comprises pores having a diameter of less than about 10 μm; and   (ii) a tumoroid population of cancerous cells and stromal cells isolated from a tumor of an animal or human, wherein the population of cells is inoculated onto said scaffold without prior incubation in a cell culture medium, wherein the tumoroid comprises a population of cells having increased resistance to a therapeutic agent compared to the population of cells isolated from the tumor of the animal or human subject but cultured in the absence of the scaffold.   
     
     
         14 . A method of generating a tumoroid comprising:
 (i) inoculating a three-dimensional scaffold of randomly oriented fibers with a population of cells removed from a tumor of an animal or human, wherein the scaffold of randomly oriented fibers are electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) and a poly(lactic-co-glycolic acid) (PLGA), and a population of cells is inoculated onto said scaffold without prior incubation of the cells in a cell culture medium; and   (ii) generating a tumoroid by incubating the inoculated scaffold in a cell culture medium under conditions allowing proliferation of the inoculated cells.   
     
     
         15 . The method of  claim 14 , wherein the scaffold of randomly oriented fibers is electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) having a ratio of PEG to PLA of from about 1:4 to about 1:10, and a poly(lactic-co-glycolic acid) (PLGA) having a lactic acid:glycolic acid ratio of about 85:15, and wherein fiber diameter ranges from about 0.3 μm to about 10 μm and the scaffold comprises pores having a diameter of less than about 10 μm. 
     
     
         16 . The method of  claim 14 , wherein the population of cells inoculated onto the scaffold comprises a mixture of cancerous cells and stromal cells. 
     
     
         17 . A method of identifying a compound that modulates the proliferation of cells of a tumor of an animal or human, the method comprising:
 (i) generating a tumoroid by:
 (a) inoculating a three-dimensional scaffold of randomly oriented fibers with a population of cells removed from a tumor of an animal or human, wherein the scaffold of randomly oriented fibers are electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) and a poly(lactic-co-glycolic acid) (PLGA), and a population of cells is inoculated onto said scaffold without prior incubation of the cells in a cell culture medium; and 
 (b) generating a tumoroid by incubating the inoculated scaffold in a cell culture medium under conditions allowing proliferation of the inoculated cells; 
   (ii) contacting the tumoroid with a compound suspected of modulating the proliferation of the cells of the tumoroid;   (iii) comparing the proliferation of the cells of the tumoroid in contact with said compound with the proliferation of the cells of a tumoroid not in contact with said compound, thereby determining if the compound modulates the proliferation of the cells of the tumoroid   
     
     
         18 . The method of  claim 17 , wherein the scaffold of randomly oriented fibers is electrospun from a mixture comprising a polyethylene glycol-polylactic acid block copolymer (PEG-PLA) having a ratio of PEG to PLA of from about 1:4 to about 1:10, and a poly(lactic-co-glycolic acid) (PLGA) having a lactic acid:glycolic acid ratio of about 85:15, and wherein fiber diameter ranges from about 0.3 μm to about 10 μm and the scaffold comprises pores having a diameter of less than about 10 μm. 
     
     
         19 . The method of  claim 17 , wherein the population of cells inoculated onto the scaffold comprises a mixture of cancerous cells and stromal cells.