Hanging drop devices, systems and/or methods

Abstract

The present disclosure relates general to devices, systems, and methods of using such devices in creating and handling hanging drops of fluid. The present disclosure also relates to cell culture devices, methods and/or systems of using such devices as well as the use of cell culture devices, for example, for research and high throughput screening.

What is claimed is: 
     
         1 .- 37 . (canceled) 
     
     
         38 . A method for co-culturing a plurality of spheroids in a single hanging drop of liquid culture media, said method comprising the steps of:
 proving an assay plate having a plurality of wells, each well extending as a hole through top and bottom surfaces of the assay plate,   filling at least one of the wells with liquid culture media so that a hanging portion of the liquid culture media hangs as a suspended drop below the well and the bottom surface of the assay plate,   culturing a first aggregated cell spheroid in the hanging portion of liquid culture media, and   inserting at least a second aggregated cell spheroid into the hanging portion of liquid culture media so that the first and second spheroids are suspended together within the same hanging portion of liquid culture media.   
     
     
         39 . The method of  claim 38 , wherein said inserting step includes extending the dispensing tip of a pipette through the well to introduce the second spheroid into the hanging portion of liquid culture media. 
     
     
         40 . The method of  claim 39 , wherein said extending step includes locating the dispensing tip into the well more than half-way between the top and bottom surfaces of the assay plate. 
     
     
         41 . The method of  claim 38 , wherein said culturing step includes locating the first spheroid entirely within the hanging portion of the liquid culture media, and said inserting step includes locating the second spheroid entirely within the hanging portion of the liquid culture media and directly adjacent the first spheroid. 
     
     
         42 . The method of  claim 38 , further including the step of surrounding the plurality of wells in the assay plate with an evaporation control liquid. 
     
     
         43 . The method of  claim 42 , wherein said surrounding step includes containing the evaporation control liquid within a water reservoir disposed along one or more edges of the assay plate. 
     
     
         44 . The method of  claim 38 , further including the step of restraining spread of the hanging portion of the liquid culture media along the bottom surface of the assay plate with a lower plateau structure surrounding each well. 
     
     
         45 . The method of  claim 38 , further including the step of restraining spread of the liquid culture media along the top surface of the assay plate with an upper plateau structure surrounding each well. 
     
     
         46 . The method of  claim 38 , further including the step of covering the top surface of the assay plate with a lid. 
     
     
         47 . The method of  claim 38 , further including the step of enclosing the bottom surface of the assay plate together with the hanging portion of the liquid culture media with a lower plate. 
     
     
         48 . The method of  claim 38 , further including the step of removing the first and second spheroids from the hanging portion of the liquid culture media through the well. 
     
     
         49 . The method of  claim 48 , wherein said removing step includes suctioning the first and second spheroids into a pipette inserted in the well. 
     
     
         50 . The method of  claim 38 , further including the step of allowing the first and second spheroids to form a co-culture within the hanging portion of the liquid culture media, and subsequently removing the co-culture through the well. 
     
     
         51 . The method of  claim 50 , wherein said removing step includes suctioning the co-culture into a pipette inserted in the well. 
     
     
         52 . A cell culturing assembly, said assembly comprising:
 an assay plate including a top surface and a bottom surface, said assay plate including a plurality of wells, each said well extending as a hole through said top and bottom surfaces of said assay plate to enable the passage of liquid culture media therethrough, each said well including a lower plateau structure extending below said bottom surface and surrounding said well,   an isolated body of liquid culture media disposed in each said well, said liquid culture media in each said well having a hanging portion suspended by surface tension from said lower plateau structure,   a plurality of cells suspended in each said body of liquid culture media and configured to aggregate into a spheroid, said spheroid disposed entirely within said hanging portion and disposed entirely below said lower plateau structure.   
     
     
         53 . The assembly of  claim 52 , wherein said top surface includes an upper reservoir configured to receive a body of liquid restricted from said plurality of wells for maintaining a substantially stable humidity above said plurality of wells. 
     
     
         54 . The assembly of  claim 53 , wherein said upper reservoir is disposed along one or more of said edges of said top surface. 
     
     
         55 . A cell culturing assembly and pipette combination, said combination comprising:
 an assay plate including a top surface and a bottom surface, said assay plate including a plurality of wells, each said well extending as a hole through said top and bottom surfaces of said assay plate to enable the passage of liquid culture media therethrough,   an isolated body of liquid culture media disposed in each said well, said liquid culture media in each said well having a hanging portion suspended by surface tension from said lower plateau structure,   a plurality of cells suspended in each said body of liquid culture media and configured to aggregate into a spheroid,   a pipette having an elongated dispensing tip, said dispensing tip having a terminal end sized with respect to said wells to extend more than half-way between said top and bottom surfaces of said assay plate.   
     
     
         56 . The combination of  claim 55 , wherein each said well has a diameter of approximately 1.6 mm. 
     
     
         57 . A method for harvesting a spheroid from a hanging drop of liquid culture media, said method comprising the steps of:
 proving an assay plate having a plurality of wells, each well extending as a hole through top and bottom surfaces of the assay plate,   filling at least one well with liquid culture media so that a portion of the liquid culture media hangs as a suspended drop below the well and the bottom surface of the assay plate,   culturing an aggregated cell spheroid in the hanging portion of liquid culture media, and   inserting the elongated dispensing tip of a pipette through the top surface opening of the one well, and suctioning the spheroid into the pipette through the dispensing tip.