Kidney organoids and methods of manufacturing
AssigneeMurdoch Childrens Research Institute
InventorsMelissa Little, Santhosh V. Kumar
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Abstract
The present disclosure relates to kidney organoids and methods of manufacturing the same. The organoids and methods may be used in a variety of applications such as disease modelling, drug screening, regenerative medicine and 5 scaling up production of kidney cells.
Claims17
An in-vitro method of producing a kidney organoid, the method comprising, swirling a population of dissociated intermediate mesoderm (IM) cells in a cell culture medium comprising fibroblast growth factor (FGF), wherein the kidney organoid comprises NPHS1+ podocytes, LTL+ proximal segments, ECAD+distal segments and ECAD+/GATA3+ collecting ducts, wherein the IM cells are swirled in culture for at least 5 days, wherein the first 24 hours comprise swirling cells in a cell culture medium comprising FGF, heparin, CHIR99021 (CHIR) and rho-associated protein kinase (ROCK) inhibitor.
The method according to 1, wherein prior to swirling the population of IM cells, the method further comprises culturing a population of stem cells for at least seven days to produce a population of IM cells, wherein the first 4 to 5 days comprise culturing stem cells in a cell culture medium comprising at least 6 μM of a Wnt/β-catenin agonist and the remaining days in culture comprises culturing cells in a cell culture medium comprising FGF and at least 0.5 μM of a Wnt/β-catenin agonist.
Dependencies are inferred from the claim text.