Three dimensional microtissue bioprinter

Abstract

A bioprinter comprises one or more dispensing units. Each dispensing unit may include (i) a syringe including a hollow body and a plunger dimensioned to translate in the body wherein the body has an exit orifice; (ii) an actuator in contact with a proximal end of the plunger; (iii) a controller for moving the actuator; and (iv) a nozzle having a wall defining a fluid path extending from an inlet of the nozzle to an outlet of the nozzle. The inlet of the nozzle is in fluid communication with the exit orifice of the syringe body. The nozzle includes a fluid passageway in fluid communication with a source of fluid and the fluid path. The bioprinter can be used in a method of preparing microtissue comprising dispensing a bioink from one or more dispensing units of the bioprinter on a plate. The microtissue may comprise cartilage cells or tumor cells or liver cells.

What is claimed is: 
     
         1 . A method of preparing microtissue, the method comprising:
 (a) providing a bioprinter comprising:
 one or more dispensing units, each dispensing unit including:
 (i) a syringe including a hollow body and a plunger dimensioned to translate in the body, the body having an exit orifice; 
 (ii) an actuator in contact with a proximal end of the plunger; 
 (iii) a controller for controlling linear motion of the actuator; and 
 (iv) a nozzle having a wall defining a fluid path extending from an inlet of the nozzle to an outlet of the nozzle, the inlet of the nozzle being in fluid communication with the exit orifice of the body of the syringe, 
 
 wherein the nozzle includes a fluid passageway in fluid communication with a source of fluid and the fluid path; and 
   (b) dispensing a bioink from one or more dispensing units of the bioprinter on a plate.   
     
     
         2 . The method of  claim 1  wherein:
 the microtissue comprises cartilage cells. 
 
     
     
         3 . The method of  claim 1  wherein:
 the microtissue comprises tumor cells, tumor stromal cells and endothelial cells. 
 
     
     
         4 . The method of  claim 1  wherein:
 the microtissue comprises liver cells. 
 
     
     
         5 . A method for repairing a bone defect or a cartilage defect, the method comprising:
 (a) providing a bioprinter comprising:
 one or more dispensing units, each dispensing unit including:
 (i) a syringe including a hollow body and a plunger dimensioned to translate in the body, the body having an exit orifice; 
 (ii) an actuator in contact with a proximal end of the plunger; 
 (iii) a controller for controlling linear motion of the actuator; and 
 (iv) a nozzle having a wall defining a fluid path extending from an inlet of the nozzle to an outlet of the nozzle, the inlet of the nozzle being in fluid communication with the exit orifice of the body of the syringe, 
 
 wherein the nozzle includes a fluid passageway in fluid communication with a source of fluid and the fluid path; 
   (b) dispensing bioink from one or more dispensing units of the bioprinter to create microtissue comprising cartilage cells; and   (c) implanting the microtissue in the bone defect or the cartilage defect.   
     
     
         6 . A method of high-throughput screening for drug discovery, the method comprising:
 (a) providing a bioprinter comprising:
 one or more dispensing units, each dispensing unit including:
 (i) a syringe including a hollow body and a plunger dimensioned to translate in the body, the body having an exit orifice; 
 (ii) an actuator in contact with a proximal end of the plunger; 
 (iii) a controller for controlling linear motion of the actuator; and 
 (iv) a nozzle having a wall defining a fluid path extending from an inlet of the nozzle to an outlet of the nozzle, the inlet of the nozzle being in fluid communication with the exit orifice of the body of the syringe, 
 
 wherein the nozzle includes a fluid passageway in fluid communication with a source of fluid and the fluid path; 
   (b) dispensing bioink from one or more dispensing units of the bioprinter to create a plurality of microtissue samples; and   (c) contacting at least a portion of the plurality of microtissue samples with a drug candidate compound.   
     
     
         7 . The method of  claim 6  wherein:
 the microtissue samples comprise tumor cells. 
 
     
     
         8 . A bioprinter comprising:
 one or more dispensing units, each dispensing unit including:
 a syringe including a hollow body and a plunger dimensioned to translate in the body, the body having an exit orifice; 
 (ii) an actuator in contact with a proximal end of the plunger; 
 (iii) a controller for moving the actuator; and 
 (iv) a nozzle having a wall defining a fluid path extending from an inlet of the nozzle to an outlet of the nozzle, the inlet of the nozzle being in fluid communication with the exit orifice of the body of the syringe, 
 wherein the syringe does not include a valve between the exit orifice of the body of the syringe and a distal end of the plunger. 
   
     
     
         9 . The bioprinter of  claim 8  wherein:
 the controller executes a program stored in the controller to drive the actuator toward the proximal end of the plunger to dispense a bioink from the body of the syringe. 
 
     
     
         10 . The bioprinter of  claim 8  further comprising:
 a temperature controller surrounding the body of the syringe. 
 
     
     
         11 . The bioprinter of  claim 8  further comprising:
 a temperature controlled plate for receiving a bioink from the outlet of the nozzle. 
 
     
     
         12 . The bioprinter of  claim 11  further comprising:
 a humidifier for creating a humidity controlled zone adjacent to the temperature controlled plate. 
 
     
     
         13 . The bioprinter of  claim 8  wherein:
 the nozzle includes a fluid passageway in fluid communication with a source of fluid and the fluid path, and 
 fluid from the fluid passageway separates droplets of dispensed material and forces the droplets of dispensed material to exit the outlet of the nozzle. 
 
     
     
         14 . The bioprinter of  claim 13  wherein:
 a volume of each droplet is in a range of 10 nanoliters to 15 microliters. 
 
     
     
         15 . The bioprinter of  claim 13  wherein:
 the source of fluid comprises a controllable valve for supplying fluid. 
 
     
     
         16 . The bioprinter of  claim 15  wherein:
 the controllable valve is controlled by a second controller. 
 
     
     
         17 . The bioprinter of  claim 13  wherein:
 the source of fluid comprises a controllable valve for supplying pulsed fluid. 
 
     
     
         18 . The bioprinter of  claim 13  wherein:
 the source of fluid comprises a controllable valve for supplying pulsed air. 
 
     
     
         19 . The bioprinter of  claim 13  wherein:
 the source of fluid comprises a controllable valve for supplying millisecond pulsed air. 
 
     
     
         20 . The bioprinter of  claim 13  wherein:
 each dispensing unit is a non-contact dispensing unit. 
 
     
     
         21 . The bioprinter of  claim 8  wherein:
 the nozzle is integral with the syringe body. 
 
     
     
         22 . The bioprinter of  claim 8  wherein:
 the nozzle is separate from the syringe body. 
 
     
     
         23 . The bioprinter of  claim 8  wherein:
 the outlet of the nozzle has an inner diameter of 100 microns to 3 millimeters. 
 
     
     
         24 . The bioprinter of  claim 8  wherein:
 the bioprinter comprises a plurality of the dispensing units. 
 
     
     
         25 . The bioprinter of  claim 8  further comprising:
 each of the dispensing units is mounted on an XYZ motion system. 
 
     
     
         26 . A method of preparing microtissue, the method comprising:
 (a) dispensing a bioink from one or more dispensing units of the bioprinter of  claim 8  on a plate.   
     
     
         27 . The method of  claim 26  wherein:
 the microtissue comprises cartilage cells. 
 
     
     
         28 . The method of  claim 26  wherein:
 the microtissue comprises tumor cells. 
 
     
     
         29 . The method of  claim 26  wherein:
 the microtissue comprises liver cells. 
 
     
     
         30 . A method for repairing a bone defect or a cartilage defect, the method comprising:
 (a) dispensing bioink from one or more dispensing units of the bioprinter of  claim 8  to create microtissue comprising cartilage cells; and   (b) implanting the microtissue in the bone defect or the cartilage defect.   
     
     
         31 . A method of high-throughput screening for drug discovery, the method comprising:
 (a) dispensing bioink from one or more dispensing units of the bioprinter of  claim 8  to create a plurality of microtissue samples; and   (b) contacting at least a portion of the plurality of microtissue samples with a drug candidate compound.   
     
     
         32 . The method of  claim 31  wherein:
 the microtissue samples comprise tumor cells.