Method and apparatus for in vitro kidney organogenesis
AssigneesThe Regents of the University of California, U.S. Department of Veterans Affairs
InventorsNorimoto Yanagawa, Masaki Nishikawa, Morgan Hamon, Peter Viktor Hauser
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Abstract
The invention provides for a device, system, and methods for using the same with kidney progenitor cells, specifically ureteric bud (UB) cells, metanephric mesenchymal (MM) cells, and the stromal cell (SC) subpopulation of the metanephric mesenchyme, to generate an embryonic kidney organoid that can be implanted into a mammalian subject to create a living, functional kidney.
Claims19
A method of generating an embryonic kidney organoid, the method comprising the steps of:
- providing a kidney mold device having a ureteric bud (UB) cell chamber and a metanephric mesenchymal (MM) cell chamber, wherein the MM cell chamber is fluidly connected to the UB cell chamber;
- providing a precursor micromold having a plurality of channels for generating a UB tube from UB cells, wherein the channels are dimensioned such that UB tubes formed in the channels fit within the UB cell chamber of the kidney mold device;
- seeding UB cells into the precursor micromold to generate a UB tube;
- transferring the UB tube into the kidney mold device UB cell chamber;
- generating a metanephric mesenchymal/stromal cell (MM/SC) aggregate or suspension;
- transferring the MM/SC aggregate or suspension into the kidney mold device MM cell chamber;
- and
- culturing the kidney mold device in growth media to generate an embryonic kidney organoid.
A method of generating an embryonic kidney organoid, the method comprising the steps of:
- providing a kidney mold device having a ureteric bud (UB) cell chamber and a metanephric mesenchymal (MM) cell chamber, wherein the MM cell chamber is fluidly connected to the UB cell chamber;
- inserting a plug into the kidney mold device MM cell chamber;
- seeding UB cells into the kidney mold device UB cell chamber to generate a UB tube;
- removing the plug by extraction or dissolution;
- generating a MM/somatic cell aggregate or suspension;
- transferring the MM/somatic cell aggregate or suspension into the kidney mold device MM cell chamber;
- and
- culturing the kidney mold device in growth media to generate an embryonic kidney organoid.
Dependencies are inferred from the claim text.