Device and method for single cell screening based on inter-cellular communication

Abstract

A method for single-cell analysis according to an embodiment of the present invention comprises: Culturing a first cell in a culture medium on a bottom side of porous membrane; Applying a sample including a second cell on a porous membrane in a culture medium; Isolating the second cell into single cell units in a pore existing in the porous membrane with a external force such as agitation and gravitational force; Generating an interaction situation between the first cells and the single cell-level second cell; Analyzing a cellular phenomena of the first cell or the second cell.

What is claimed is: 
     
       1. A device for single-cell analysis comprising:
 a substrate; 
 a culture medium disposed on the substrate, the culture medium includes a plurality of first cells cultured therein; 
 a porous membrane disposed on the culture medium, the porous membrane has a plurality of pores capable of isolating a cell into single cell units; and 
 a plurality of second cells isolated in some of the pores of the porous membrane, 
 wherein a gap is formed between the substrate and the porous membrane which is a space for the culture medium, 
 wherein the gap continues throughout below the porous membrane, 
 wherein each of the pores penetrates the porous membrane from top surface to bottom surface, 
 wherein a diameter of the pore is 1 to 100 μm, and wherein the first cell is a fibroblast cell and the second cell is a tumor cell. 
 
     
     
       2. The device of  claim 1 ,
 wherein the gap between the porous membrane and the substrate is 1 to 100 μm. 
 
     
     
       3. The device of  claim 1 ,
 wherein the porous membrane is made of a material selected from polymeric or inorganic materials. 
 
     
     
       4. The device of  claim 3 ,
 wherein the porous membrane is made of a photosensitive polymeric material. 
 
     
     
       5. The device of  claim 4 ,
 wherein the porous membrane is made by forming a pore in a photosensitive polymeric membrane through a lithography method. 
 
     
     
       6. The device of  claim 3 ,
 wherein the porous membrane is made by forming a pore in a polymeric membrane through a soft lithography method. 
 
     
     
       7. The device of  claim 1 ,
 wherein the porous membrane has pores of 10 2  to 10 6  holes/cm 2 . 
 
     
     
       8. The device of  claim 1 ,
 wherein the porous membrane has pores and an interval between the pores is 1 μm to 10 mm. 
 
     
     
       9. A method for single-cell analysis using the device of  claim 1  comprising:
 Culturing a first cell in a culture medium on a bottom side of porous membrane; Applying a sample including a second cell on a porous membrane in a culture medium; Isolating the second cell into single cell units in a pore existing in the porous membrane with an external force such as agitation and gravitational force; Generating an interaction situation between the first cells and the single cell-level second cell; Analyzing cellular phenomena of the first cell or the second cell. 
 
     
     
       10. The method of  claim 9 ,
 wherein the first cell is a fibroblast cell and the second cell is a tumor cell. 
 
     
     
       11. The method of  claim 9 ,
 wherein a thickness of the culture medium is 1 to 100 μm. 
 
     
     
       12. The method of  claim 9 ,
 wherein a concentration of the first cell is 1×10 5  to 1×10 7  cells/mL. 
 
     
     
       13. The method of  claim 9 ,
 wherein when applying the second cells, stirring is performed at the same time. 
 
     
     
       14. The method of  claim 13 ,
 wherein the stirring is performed for 1 minute to 1 hour at 10 to 500 rpm. 
 
     
     
       15. The method of  claim 9 ,
 wherein a concentration of the second cell in the sample i (a number of pores in a porous membrane×1) to (a number of pores in a porous membrane×10,000) cells/mL or 1×10 2  to 1×10 10  cells/mL. 
 
     
     
       16. The method of  claim 9 ,
 wherein a diameter of the pore is 1 to 100 μm. 
 
     
     
       17. The method of  claim 9 ,
 wherein the porous membrane has pores of 10 2  to 10 6  holes/cm 2 . 
 
     
     
       18. The method of  claim 9 ,
 wherein the porous membrane has pores and a gap between the pores is 1 μm to 10 mm. 
 
     
     
       19. The method of  claim 9 ,
 wherein the interaction is generated by contact or paracrine factors between the first cell and the second cell for 1 hour to 7 days. 
 
     
     
       20. The method of  claim 9 ,
 wherein the analyzing a cell activity of the first cell or the second cell further comprises screening the cell activity of the first cell or the second cell. 
 
     
     
       21. The method of  claim 9 ,
 wherein the analyzing a cell activity of the first cell or the second cell further comprises capturing and analyzing the second cell.