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renal progenitor cells

A progenitor cell and cell technology, applied in the field of kidney development, can solve problems such as lack of understanding and hindering the realization of human pluripotent stem cells

Active Publication Date: 2021-10-26
THE UNIV OF QUEENSLAND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, a lack of understanding of how to produce the essential cell types that give rise to nephrons and other structures of the kidney has hampered the realization of human pluripotent stem cells as a source of cells for clinical use and as therapeutic agents for diseases such as kidney

Method used

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Embodiment

[0152] Materials and methods

[0153] hESC culture and differentiation

[0154] In supplemented with 20% KnockOut Serum Replacement (Life Technologies), 100 μM MEM NEAA (Life Technologies), 110 μM 2-Mercaptoethanol (Life Technologies), 1x Penicillin / Streptomycin (Life Technologies), 1x Glutamax (Life Technologies) and 10 ng / HES3 (MIXL1 GFP / wt )cell. The day before initiation of differentiation, cells were incubated at 12,000-15,000 cells / cm 2 Plate on Matrigel coated 96-well plates. After overnight culture, cells were exposed to 30ng / mL BMP4 (R&Dsystems) and 10ng / mL Activin A (R&Dsystems) or 8μM CHIR99021 in previously established serum-free medium APEL for 2-3 days, and then exposed to APEL culture 200 ng / mL FGF9 and 1 μg / mL heparin in the medium were maintained for 4 days to induce IM cells. In the case of BMP4 / Activin A induction, cells were subsequently exposed to 200 ng / mL FGF9, 50 ng / mL BMP7, 0.1 μM RA and 1 μg / mL heparin for 4-11 days. In the case of CHIR99021 ...

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Abstract

There is provided a method for the simultaneous production of nephron progenitor cells and ureteric epithelial progenitor cells, comprising the step of contacting intermediate mesoderm cells with fibroblast growth factor 9 and / or fibroblast growth factor 20 , and optionally one or more selected from: bone morphogenetic protein 7; heparin; Wnt agonists; retinoic acid; The concentrations of Wnt agonist, retinoic acid and / or RA antagonist can be manipulated to favor the relative production of nephron progenitor cells and ureteric epithelial progenitor cells. The intermediate mesoderm cells are ultimately derived from human pluripotent stem cells via the posterior primitive streak stage. The nephron progenitor cells and ureteric epithelial progenitor cells may have end uses such as for kidney repair and regeneration, bioprinting of kidneys, and screening of compounds for nephrotoxicity.

Description

technical field [0001] The present invention relates to kidney development. More specifically, the present invention relates to in vitro methods for the production of nephron progenitor cells and ultimately ureteral progenitor cells from human pluripotent stem cells. Background technique [0002] As prevalence of end-stage kidney disease rises to 8% globally 1 , an urgent need for renal regeneration strategies. The kidney is a mesoderm organ that differentiates from intermediate mesoderm (IM) through the formation of the ureteric bud (UB) and the interaction between this bud and the adjacent IM-derived metanephric mesenchyme (MM) 2 . Nephrons are derived from a population of nephron progenitor cells derived from MM 3 . IM itself is derived from the back original article 4 . Although the developmental origins of the kidney are well understood 2 , however nephron formation in the human kidney is complete before birth 5 . Therefore, there are no postnatal stem cells c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/0735C12N5/074A61K35/22A61P13/12C12Q1/02
CPCC12N2501/119C12N2501/155C12N2501/16C12N2501/385C12N2501/415C12N2501/91C12N2506/02C12N2506/45C12N5/0687A61P13/12G01N33/5014G01N33/5044
Inventor 梅利莎·利特尔高里实
Owner THE UNIV OF QUEENSLAND
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