Microchip high density hanging drop three-dimension culture platform
AssigneesThe Board of Trustees of the University of Illinois, Mayo Foundation for Medical Education and Research
InventorsAnurup GANGULI, Rashid BASHIR, Panagiotis Z. ANASTASIADIS, George VASMATZIS
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Abstract
Provided are methods and related devices for preparing a cell and tissue culture, including a hanging drop culture. Microwells are specially loaded with cell mixtures using a removable reservoir and forcing cells into the underlying microwells. The removable reservoir is removed and the cells partitioned into the individual microwells and covered by an immiscible layer of fluid. The microwells and immiscible layer is inverted and the cells in the microwells cultured. The microwells may have shape-controlling elements to control the three-dimensional shape of the culture.
Claims20
A method for preparing a cell and tissue culture, the method comprising the steps of:
- providing an array of surface-oxidized, hydrophilic microwells, wherein each microwell has:
- a depth of between 50 μm to 1000 μm;
- a longest dimension of between 50 μm to 3000 μm;
- a separation distance from an adjacent microwell that prevents unwanted liquid leaking between adjacent microwells;
- forming a removable reservoir over at least a portion of a top surface of the microwells;
- loading a mixture comprising cells in the removable reservoir and forcing the mixture comprising cells in the removable reservoir into the microwells;
- removing the removable reservoir from the microwells;
- covering the microwells with the cells with an immiscible layer;
- partitioning the cells into the individual microwells;
- inverting the array of microwells with the cells and the immiscible layer so that the immiscible layer confines the cells to the individual microwells;
- and
- culturing the cells in the inverted array of microwells, thereby preparing the cell and tissue culture.
An array of microwells with fluid exchange for cell and tissue culture comprising:
- a substrate comprising an array of microwells wherein each microwell has a depth of between 80 μm to 160 μm and a longest dimension of between 100 to 500 μm;
- fluidic channels having a width of 5 to 15 μm that fluidly interconnect the microwells such that each individual microwell in the array is fluidly connected to an adjacent microwell;
- and
- at least one cell and tissue culture media reservoir well having a larger fluid capacity than the array of microwells, wherein said cell and tissue culture media reservoir well is fluidly connected to and positioned peripherally relative to the array of microwells;
- wherein the surface of the array of microwells are oxidized and surface treated to improve hydrophilicity and the substrate comprises silicon, oxide, glass or plastic.
Dependencies are inferred from the claim text.