Method for inducing differentiation of intermediate mesodermal cell to renal progenitor cell, and method for inducing differentiation of pluripotent stem cell to renal progenitor cell

A technology of pluripotent stem cells and cell induction, which is applied in the field of induction of renal progenitor cells from differentiation of intermediate mesoderm cells and induction of renal progenitor cells from differentiation of pluripotent stem cells.

Pending Publication Date: 2020-01-07
KYOTO UNIV
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Problems solved by technology

[0005] Several methods for inducing nephron progenitor cells from human iPS cells and human ES cells have been reported (non-patent documents 3 to 6), but there are low differentiation induction efficiency due to the use of embryoid bodies (embryoid bodies, EBs) (non-patent documents 3-6). Patent Document 3), the problem that it is impossible to determine whether each stage of occurrence has been correctly reproduced (Non-Patent Documents 4 to 6)

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  • Method for inducing differentiation of intermediate mesodermal cell to renal progenitor cell, and method for inducing differentiation of pluripotent stem cell to renal progenitor cell
  • Method for inducing differentiation of intermediate mesodermal cell to renal progenitor cell, and method for inducing differentiation of pluripotent stem cell to renal progenitor cell
  • Method for inducing differentiation of intermediate mesodermal cell to renal progenitor cell, and method for inducing differentiation of pluripotent stem cell to renal progenitor cell

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Embodiment

[0139] Hereinafter, although this invention is demonstrated concretely based on an Example, this invention is not limited to the following aspect.

[0140] Induction of renal progenitor cells from iPS cell differentiation

[0141] Kidney progenitor cells were induced from iPS cell differentiation according to the following protocol. Among them, 201B7-derived OSR1-GFP / SIX2-tdTomato reporter-human iPS cells were used as iPS cells.

[0142] 1. Undifferentiated iPS cells were single-celled by accutase treatment, suspended in ReproFF2 medium (ReproCELL) supplemented with 10 μM Y27632, and placed in a well (well) coated with Matrigel (BD Biosciences) at 1.0×10 4 / well~5.0×10 4 Sow at a density of / well and incubate at 37°C for 24 hours.

[0143] 2. After 24 hours (day 1), use DMEM / F12 Glutamax (Thermo Fisher Scientific Inc.) supplemented with vitamin A free B27 supplement (Thermo Fisher Scientific Inc.) The culture medium of xanthic acid, 1ng / ml BMP4, and 100ng / ml FGF2 was used...

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Abstract

A method for producing renal progenitor cells from pluripotent stem cells, comprising steps (i) to (vi): (i) culturing the pluripotent stem cells in a culture medium containing FGF2, BMP4, a GSK-3[beta] inhibitor and retinoic acid or a derivative thereof; (ii) culturing cells produced in step (i) in a culture medium containing FGF2, a GSK-3[beta] inhibitor and BMP7; (iii) culturing cells producedin step (ii) in a culture medium containing FGF2, a GSK-3[beta] inhibitor, BMP7 and a TGF[beta] inhibitor; (iv) culturing cells produced in step (iii) in a culture medium containing FGF2, a GSK-3[beta] inhibitor, BMP7, activin and a ROCK (Rho-kinase) inhibitor; (v) culturing cells produced in step (iv) in a culture medium containing retinoic acid or a derivative thereof and FGF9; and (vi) culturing cells produced in step (v) in a culture medium containing a GSK-3[beta] inhibitor and FGF9 to induce renal progenitor cells from intermediate mesodermal cells.

Description

technical field [0001] The present invention relates to a method for inducing renal progenitor cells differentiated from intermediate mesoderm cells. The present invention also relates to a method for inducing renal progenitor cells differentiated from pluripotent stem cells. Background technique [0002] Currently, the number of chronic kidney disease (CKD) patients in Japan is estimated to be about 13 million, and it is called a new national disease. There are few radical treatments for chronic kidney disease. According to its progress, there are more than 300,000 patients with end-stage chronic renal failure who must undergo dialysis treatment. This is not only a major medical problem, but also a major medical economic problem. question. As a radical cure for chronic kidney disease including end-stage chronic renal failure, kidney transplantation can be cited, but the supply has not kept pace with the demand due to a serious shortage of donor organs. [0003] The kidne...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0735A61K35/22A61L27/38A61P13/12C12N5/071C12N5/10C12N15/09
CPCA61K35/22A61P13/12C12N15/09C12N5/0687C12N5/0068C12N2533/52C12N2501/115C12N2501/15C12N2501/155C12N2501/727C12N2501/998C12N2506/45A61L27/3895C12N5/0686C12N5/0662A61L27/38C12N2500/30C12N2501/119C12N2501/16C12N5/0696
Inventor 末田伸一笠原朋子长船健二
Owner KYOTO UNIV
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