Method of Growing Cells on a Droplet Actuator

Abstract

A method of growing cells on a droplet actuator is provided. The method may include providing a droplet actuator including a cell culture droplet including a cell culture medium and one or more cells; and maintaining the droplet at a temperature suitable for causing the cells to grow in the cell culture medium on the droplet actuator. Related methods, droplet actuators, and systems are also provided.

1 - 4 . (canceled) 
     
     
         5 . A method of growing cells on a droplet actuator, the method comprising the steps:
 a. providing a droplet actuator comprising a sample droplet loaded thereon and a cell culture medium, wherein the sample droplet comprises one or more cells and is in an oil filler fluid;   b. merging the sample droplet and the cell culture medium by electrowetting to form a cell culture droplet in the oil filler fluid and comprising the one or more cells and cell culture medium; and   c. maintaining the cell culture droplet in the oil filler fluid at a temperature suitable for causing the cells to grow in the cell culture medium on the droplet actuator.   
     
     
         6 . The method of claim  1  wherein the cell culture droplet is situated between droplet actuator substrates in proximity to a droplet operations surface. 
     
     
         7 . The method of claim  1  wherein the one or more cells are bound to beads. 
     
     
         8 . The method of claim  1 , wherein the sample droplet and cell culture droplet are substantially surrounded by the oil filler fluid. 
     
     
         9 . The method of claim  1 , wherein the oil comprises silicone oil. 
     
     
         10 . The method of claim  1 , wherein the one or more cells of the cell culture droplet comprises only one cell. 
     
     
         11 . The method of claim  1 , wherein the droplet actuator comprises a heating element configured to heat the cell culture medium to an appropriate temperature for incubation. 
     
     
         12 . The method of claim  1 , wherein the droplet actuator is associated with a heating element configured to heat the cell culture medium to an appropriate temperature for incubation. 
     
     
         13 . The method of claim  1 , further comprising a step prior to step (b) wherein, a cell sorting process is conducted on the droplet actuator to produce the droplet comprising one or more cells, and wherein the droplet comprising one or more cells comprises one or more cells of the same cell type. 
     
     
         14 . The method of claim  1 , wherein the cell culture medium is in a cell culture reservoir on the droplet actuator. 
     
     
         15 . The method of claim  1 , wherein the cell culture medium is in contact with the atmosphere or a sub-atmosphere on the droplet actuator. 
     
     
         16 . The method of claim  1 , wherein electrowetting electrodes fluidically connect the cell culture droplet with a fluid reservoir. 
     
     
         17 . The method of  claim 16 , wherein the fluid reservoir comprises a volume of reagent fluid. 
     
     
         18 . The method of  claim 17 , wherein after step  1 ( b ), droplet operations are conducted to merge the cell culture droplet with a droplet comprising the reagent fluid from the fluid reservoir, wherein the reagent fluid is exchanged with the cell culture medium in the cell culture droplet. 
     
     
         19 . The method of claim  1 , wherein droplet operations are conducted to merge the cell culture droplet with a droplet comprising beads. 
     
     
         20 . The method of  claim 19 , wherein the beads have an affinity for one or more cells in the cell culture droplet. 
     
     
         21 . The method of  claim 20 , wherein the one or more cells are bound to the beads in the cell culture droplet. 
     
     
         22 . The method of  claim 21 , wherein the beads are immobilized within the cell culture droplet. 
     
     
         23 . The method of  claim 22 , wherein the beads are magnetically responsive beads, and wherein the magnetically responsive beads are magnetically immobilized within the cell culture droplet. 
     
     
         24 . The method of  claim 22 , wherein the beads are immobilized within the cell culture droplet by one or more physical barriers. 
     
     
         25 . The method of  claim 22 , wherein droplet operations are conducted to split the cell culture droplet to form one or more droplets substantially lacking beads, wherein the one or more droplets substantially lacking beads comprise reagent fluid and cell culture medium, and wherein the one or more droplets substantially lacking beads are transported away from the cell culture droplet. 
     
     
         26 . The method of  claim 25 , wherein the merging and splitting droplet operations deliver a metabolically useful substance, drug, or chemical to the cell culture medium of the cell culture droplet. 
     
     
         27 . The method of  claim 25 , wherein the merging and splitting droplet operations change the pH concentration of the cell culture medium of the cell culture droplet. 
     
     
         28 . The method of  claim 18 , wherein droplet operations are conducted to split the cell culture droplet to form one or more droplets, wherein the merging and splitting droplet operations change the concentration of cells within the cell culture medium of the cell culture droplet.