A method is provided for producing renal organoids comprising nephrons, ureteric bud and vasculature and/or progenitors of these. In one embodiment, the methods includes contacting intermediate mesoderm cells with: fibroblast growth factor 9 and/or fibroblast growth factor 20 and/or fibroblast growth factor 2 and optionally, one or more selected from the group consisting of: bone morphogenic protein 7; heparin; a Wnt agonist; retinoic acid; and an RA antagonist under conditions that promote formation of vascularized renal organoids. Another embodiment includes producing mesoderm cells by sequentially contacting pluripotent stem cells with a Wnt agonist and fibroblast growth factor 9 and/or fibroblast growth factor 20 and/or fibroblast growth factor 2, followed by a relatively short re-exposure to the Wnt agonist. The renal organoids may have end uses such as for kidney repair and regeneration, bioprinting of kidneys or functional components thereof, renal cell arrays and screening compounds for nephrotoxicity.

A medium for culturing and expanding nephron progenitor cells, the medium containing a GSK-3 inhibitor, a ROCK inhibitor, and at least one fibroblast growth factor selected from the group consisting of FGF9 and FGF20. Also, a method for culturing and expanding nephron progenitor cells using the medium. Also, a method for producing renal organoids, the method including a step of culturing and expanding nephron progenitor cells using the above method for culturing and expanding nephron progenitor cells, and a step of differentiating the cultured and expanded nephron progenitor cells into renal organoids.

A method of producing renal progenitor cells from pluripotent stem cells involves culturing pluripotent stem cells in a medium containing FGF2, BMP4, a GSK-3 inhibitor, and retinoic acid or a derivative thereof, culturing the resulting cells in a medium containing FGF2, a GSK-3 inhibitor, and BMP7, culturing the resulting cells in a medium containing FGF2, a GSK-3 inhibitor, BMP7, and a TGF inhibitor, culturing the resulting cells in a medium containing FGF2, a GSK-3 inhibitor, BMP7, activin, and a ROCK (Rho-kinase) inhibitor, culturing the resulting cells in a medium containing retinoic acid or a derivative thereof, and FGF9, and culturing the resulting cells in a medium containing a GSK-3 inhibitor and FGF9, to induce renal progenitor cells from intermediate mesodermal cells.

A method for acquiring and producing high-purity renal progenitor cells from a renal progenitor cell population into which pluripotent stem cells are induced to differentiate, by identifying a cell surface antigen marker specific to renal progenitor cells. The disclosed method may include, for example, the steps of: (i) culturing the pluripotent stem cells under conditions that induce differentiation into renal progenitor cells; and (ii) sorting a cell population from the cells obtained at step (i), by using at least one cell surface marker selected from the group consisting of CD9(), CD55(), CD106(+), CD140a(+), CD140b(+), CD165(+), CD271(+) and CD326().

The following disclosure generally relates to methods of culturing podocytes in vitro. The cultures can be used for drug screening (such as medium or high throughput drug screening), and for studying molecular pathways involved in glomerular diseases. The disclosure also provides methods for analyzing the healthiness of podocytes in a cell culture. The disclosure also relates to diagnosis of kidney diseases.

The present disclosure provides compositions of matter, methods and instruments for nucleic acid-guided nickase/reverse transcriptase fusion enzyme editing of nucleic acids in live mammalian cells.

The disclosure describes novel systems, methods, and compositions for the manipulation of nucleic acids in a targeted fashion. The disclosure describes non-naturally occurring, engineered CRISPR systems, components, and methods for targeted modification of nucleic acids. Each system includes one or more protein components and one or more nucleic acid components that together target nucleic acids.

A method of producing a mesodermal-lineage primitive streak cell includes a first step of culturing pluripotent stem cells in a medium containing FGF2, BMP (bone morphogenetic protein) 4, a GSK-3 inhibitor, and retinoic acid or a derivative thereof and a second step of culturing cells obtained in the first step in a medium containing FGF2, a GSK-3 inhibitor, and BMP7.

The present disclosure provides compositions of matter, methods and instruments for nucleic acid-guided nickase/reverse transcriptase fusion enzyme editing of nucleic acids in live mammalian cells.

Isolated or purified mammalian kidney-derived cell populations from mammalian kidney tissue are provided. Methods are provided for the isolation and purification of the mammalian kidney-derived cell population. Methods for treating kidney disease are provided by administration of the isolated or purified mammalian kidney-derived cell population to a mammalian subject.