Organoids in extracellular matrix droplets
AssigneePresident and Fellows of Harvard College
InventorsDavid A. Weitz, Iris Elke Ruider, Jean Carlos Serrano Flores, Andreas R. Bausch, Perry Ellis
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Abstract
Methods, articles, and fluidic systems for sorting and growing organoids are generally provided. Organoids may be useful for various applications, such as certain types of biological studies, or for testing the biocompatibility and performance of drugs. However, conventional methods often struggle to identify, isolate, and use organoids. In some embodiments, using the droplet-based methods and associated articles and fluidic systems provided herein, organoids may be selected, isolated, and studied.
Claims30
A method, comprising:
- growing a plurality of organoids in a first plurality of droplets;
- and
- sorting the first plurality of droplets to produce a second plurality of droplets wherein greater than or equal to 50% of the droplets of the second plurality of droplets comprise organoids.
A method, comprising:
- exposing a plurality of droplets containing organoids to an agent suspected of interacting with the organoids;
- and
- determining an effect of the agent on the organoids.
The method of any one of the preceding claims, wherein at least 50% of the organoids have a luminal structure.
The method of any one of the preceding claims, wherein at least 50% of the organoids have a ratio of real volume to reduced volume that is greater than or equal to 125%.
The method of any one of the preceding claims, wherein greater than or equal to 50% of the organoids are branched organoids having a maximum tip-to-tip pathlength of at least 100 microns.
The method of any one of the preceding claims, wherein at least 50% of the organoids have a maximum tip-to-tip pathlength that is greater than or equal to 1.5× a cube-root of a real volume of the organoid.
The method of any one of the preceding claims, wherein greater than or equal to 50% of the organoids are branched organoids.
The method of any one of the preceding claims, wherein the maximum tip to tip path length is greater than or equal to 125 microns.
The method of any one of the preceding claims, wherein the organoid comprises cancer cells.
The method of any one of the preceding claims, wherein the organoid comprises human cells.
The method of any one of the preceding claims, wherein the method comprises growing greater than or equal to 50% of the organoids from single cells.
The method of any one of the preceding claims, wherein the sorting is performed at least in part by imaging at least some droplets of the plurality of droplets.
The method of any one of the preceding claims, wherein the sorting is performed at least in part by measuring an optical property of at least some droplets of the plurality of droplets.
The method of any one of the preceding claims, wherein the sorting of the organoids is at least partially based on a presence or absence of a particular gene or protein of the organoid, the presence or absence of a particular morphological feature of the organoid, and/or a distribution of a protein within the organoid.
The method of any one of the preceding claims, further comprising sorting the first plurality of droplets to produce a third plurality of droplets wherein greater than or equal to 50% of the droplets of the third plurality of droplets comprise organoids, wherein at least 50% of the organoids of the third plurality of droplets differ from at least 50% of the organoids of the second plurality of droplets based on a difference in the presence or absence of a particular gene or protein or the presence or absence of a particular morphological feature.
A method, comprising:
- containing an organoid in a droplet containing a gel-forming solution;
- gelling the gel-forming solution;
- and
- growing the organoid to a size exceeding the average diameter of the droplet.
An article, comprising:
- a plurality of cell clusters in a gel, wherein at least 50% of the cell clusters are organoids.
An article comprising:
- a plurality of droplets, wherein greater than or equal to 50% of the droplets of the plurality of droplets comprise an organoid.
Dependencies are inferred from the claim text.