High-throughput culture and transfer device and method

Abstract

A method and device for creating hanging drop cell aggregates. The method and device includes a plurality of pegs that allow for high throughput culture of aggregates. Also disclosed are means of transferring formed aggregates to various destinations, such as well plates, scaffolding, tissues, or wounds. Use of the device permits aggregates to be prepared or created in larger quantities than current methods, and allows for them to be transferred more efficiently.

1 .- 38 . (canceled) 
     
     
         39 . A system for creating cell aggregates, said system comprising:
 a plurality of pegs of predetermined size and arranged in a predetermined configuration; and   said plurality of pegs are configured for hanging said applied cells from said plurality of pegs, wherein said hanging comprises an inversion of said plurality of pegs, to provide the creation of the hanging drop cell aggregates.   
     
     
         40 . The system of  claim 39 , further comprising:
 an incubation chamber to accommodate said plurality of pegs containing said aggregates.   
     
     
         41 . The system of  claim 40 , further comprising:
 said incubation chamber configured to accommodate said plurality of pegs in an inverted position.   
     
     
         42 . The system of  claim 41 , wherein said incubation chamber is configured to hold an incubation medium. 
     
     
         43 . The system of  claim 42  wherein said incubation medium comprises at least one of:
 phosphate buffer solution, saline, growth medium, basal growth medium, any commercially available medium, or custom prepared medium. 
 
     
     
         44 . (canceled) 
     
     
         45 . The system of  claim 39 , further comprising:
 a well plate, wherein said well plate comprises a plurality of wells that are spaced and sized in accordance with said plurality of said pegs having said aggregates; and   wherein said well plate is configured whereby said aggregates can be transferred from said plurality of pegs to respective said plurality of wells of said well plate by positioning said plurality of pegs to coincide with said plurality of wells.   
     
     
         46 . The system of  claim 45 , wherein the transferred cells are used for at least one of the following: further culture, maintenance, study, research, characterization, or some other purpose. 
     
     
         47 . The system of  claim 45 , wherein:
 said wells in said plurality of wells contain a medium; and   wherein said well plate is configured wherein said aggregates contact with said medium to accomplish said transfer.   
     
     
         48 . The system of  claim 47 , wherein said medium comprises at least one of the following: growth medium, DMEM F-12 10% calf serum medium, any commercially available medium, or custom prepared medium. 
     
     
         49 . The system of  claim 45 , further comprising:
 forced air, wherein said forced air is applied to said aggregates to accomplish said transfer of said aggregates.   
     
     
         50 . The system of  claim 49 , wherein said forced air is applied through and/or adjacent to said plurality of pegs. 
     
     
         51 . The system of  claim 45 , further comprising:
 a forced medium source, wherein said forced medium source applies said forced medium to said aggregates to accomplish said transfer of said aggregates.   
     
     
         52 . The system of  claim 45 , further comprising:
 a pusher means for pushing said aggregates to accomplish said transfer, said pushing being of sufficient force to dislodge said aggregates.   
     
     
         53 . The system of  claim 45 , further comprising:
 a plunger configured to push said aggregates to accomplish said transfer, wherein said pushing being of sufficient force to dislodge said aggregates.   
     
     
         54 . The system of  claim 45 , further comprising:
 an agitation means for agitating said aggregates to accomplish said transfer, said agitation being of sufficient force to dislodge said aggregates.   
     
     
         55 . The system of  claim 45 , further comprising:
 an agitator device in communication with said culture plate, said agitator device providing sufficient agitation force to dislodge said aggregates.   
     
     
         56 . The system of  claim 45 , wherein said plurality of pegs is disposed on a culture plate, and said system further comprising a guide plate, wherein said guide plate is configured to receive said culture plate to effect said positioning. 
     
     
         57 . The system of  claim 39 , further comprising:
 a scaffolding, said scaffolding comprising a plurality of apertures spaced and sized in accordance with the plurality of said pegs having said aggregates; and   wherein said scaffolding is configured whereby said aggregates can be transferred from said plurality of pegs to respective said plurality of apertures in said scaffolding by positioning said plurality of pegs to coincide with said plurality of apertures.   
     
     
         58 . The system of  claim 57 , wherein said apertures comprise at least one of; holes, target sites, indententations or pores or any combination thereof. 
     
     
         59 . The system of  claim 57 , wherein said scaffolding comprises at least one of:
 predetermined destination object, bandage, sponge, tissue-engineered construct, biomaterial construct or a combination thereof.   
     
     
         60 . The system of  claim 57 , further comprising:
 forced air, wherein said forced air is applied to said aggregates to accomplish said transfer of said aggregates.   
     
     
         61 . The system of  claim 60 , wherein forced air is applied through and/or adjacent to said plurality of pegs. 
     
     
         62 . The system of  claim 57 , further comprising:
 a pusher means for pushing said aggregates to accomplish said transfer, said pushing being of sufficient force to dislodge said aggregates.   
     
     
         63 . The system of  claim 57 , further comprising:
 an agitation means for agitating said aggregates to accomplish said transfer, said agitation of sufficient force to dislodge said aggregates.   
     
     
         64 . The method of  claim 39 , further comprising:
 a scaffolding;   a punching means for forming a plurality of apertures spaced and sized in accordance with the plurality of said pegs having said aggregates; and   wherein said scaffolding is formed by said punching means whereby said aggregates can be transferred from said plurality of pegs to respective said plurality of apertures in said scaffolding by positioning said plurality of pegs to coincide with said plurality of apertures.   
     
     
         65 . The system of  claim 64 , wherein said apertures comprise at least one of; holes, target sites, indententations or pores or any combination thereof. 
     
     
         66 . The system of  claim 64 , further comprising:
 forced air, wherein said forced air is applied to said aggregates to accomplish said transfer of said aggregates.   
     
     
         67 . The system of  claim 66 , wherein forced air is applied through and/or adjacent to said plurality of pegs. 
     
     
         68 . The system of  claim 64 , further comprising:
 a pusher means for pushing said aggregates to accomplish said transfer, said pushing being of sufficient force to dislodge said aggregates.   
     
     
         69 . The system of  claim 64 , further comprising:
 an agitation means for agitating said aggregates to accomplish said transfer, said agitation of sufficient force to dislodge said aggregates.   
     
     
         70 . The system of  claim 64 , wherein said scaffolding comprises at least one of:
 predetermined destination object, bandage, sponge, tissue-engineered construct, biomaterial construct, synthetic matrix protein, organic matrix protein, extracellular matrix protein, or a combination thereof.   
     
     
         71 . The system of  claim 39 , wherein said plurality of pegs are configured to allow said aggregates to be transferred to the wound or other tissue bed of a subject. 
     
     
         72 . The system of  claim 71 , wherein the wound or other tissue bed of a subject comprises at least one of: heart tissue, skeletal muscle tissue, fat tissue, brain tissue, skin, or other tissues. 
     
     
         73 . The system of  claim 39 , wherein said aggregates comprise at least one of:
 human adipose-derived stromal cell multicellular aggregates, tumor cell aggregates, mammospheres, stem cell aggregates, hybrid cell aggregates or any and all cell types and combinations thereof.   
     
     
         74 . The system of  claim 39 , wherein at least some of each of said plurality of pegs are between about 1 mm and about 5 mm in diameter. 
     
     
         75 . The system of  claim 39 , wherein said at least some of each of said plurality of pegs are less than about 1 mm in diameter. 
     
     
         76 . The system of  claim 39 , wherein at least some of each of said plurality of pegs are greater than about 5 mm in diameter. 
     
     
         77 . The system of  claim 39 , wherein said plurality of pegs numbers greater than 1. 
     
     
         78 . The system of  claim 39 , wherein said plurality of pegs numbers greater than about 50. 
     
     
         79 . The system of  claim 39 , wherein said plurality of pegs numbers greater than about 100.