دورية أكاديمية
A Toolbox to Characterize Human Induced Pluripotent Stem Cell-Derived Kidney Cell Types and Organoids.
| العنوان: | A Toolbox to Characterize Human Induced Pluripotent Stem Cell-Derived Kidney Cell Types and Organoids. |
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| المؤلفون: | Vanslambrouck JM; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Wilson SB; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Tan KS; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Soo JY; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Scurr M; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Spijker HS; Murdoch Children's Research Institute, Melbourne, Victoria, Australia.; Department of Nephrology, Leiden University Medical Center, Leiden, The Netherlands., Starks LT; Murdoch Children's Research Institute, Melbourne, Victoria, Australia., Neilson A; Department of Genetics, Genome Engineering and iPSC Center and., Cui X; Department of Genetics, Genome Engineering and iPSC Center and., Jain S; Department of Medicine, Kidney Translational Research Center, Washington University School of Medicine, St. Louis, Missouri; and., Little MH; Murdoch Children's Research Institute, Melbourne, Victoria, Australia; melissa.little@mcri.edu.au.; Faculty of Medicine, Dentistry and Health Sciences, Department of Paediatrics and.; Department of Anatomy and Neuroscience, University of Melbourne, Victoria, Australia., Howden SE; Murdoch Children's Research Institute, Melbourne, Victoria, Australia.; Faculty of Medicine, Dentistry and Health Sciences, Department of Paediatrics and. |
| المصدر: | Journal of the American Society of Nephrology : JASN [J Am Soc Nephrol] 2019 Oct; Vol. 30 (10), pp. 1811-1823. Date of Electronic Publication: 2019 Sep 06. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Wolters Kluwer Health, on behalf of the American Society of Nephrology Country of Publication: United States NLM ID: 9013836 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1533-3450 (Electronic) Linking ISSN: 10466673 NLM ISO Abbreviation: J Am Soc Nephrol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2023- : Hagerstown, MD : Wolters Kluwer Health, on behalf of the American Society of Nephrology Original Publication: Baltimore, MD : Williams & Wilkins, c1990- |
| مواضيع طبية MeSH: | Induced Pluripotent Stem Cells* , Organoids*, Kidney/*cytology, Animals ; Female ; Mice ; Organogenesis |
| مستخلص: | Background: The generation of reporter lines for cell identity, lineage, and physiologic state has provided a powerful tool in advancing the dissection of mouse kidney morphogenesis at a molecular level. Although use of this approach is not an option for studying human development in vivo , its application in human induced pluripotent stem cells (iPSCs) is now feasible. Methods: We used CRISPR/Cas9 gene editing to generate ten fluorescence reporter iPSC lines designed to identify nephron progenitors, podocytes, proximal and distal nephron, and ureteric epithelium. Directed differentiation to kidney organoids was performed according to published protocols. Using immunofluorescence and live confocal microscopy, flow cytometry, and cell sorting techniques, we investigated organoid patterning and reporter expression characteristics. Results: Each iPSC reporter line formed well patterned kidney organoids. All reporter lines showed congruence of endogenous gene and protein expression, enabling isolation and characterization of kidney cell types of interest. We also demonstrated successful application of reporter lines for time-lapse imaging and mouse transplantation experiments. Conclusions: We generated, validated, and applied a suite of fluorescence iPSC reporter lines for the study of morphogenesis within human kidney organoids. This fluorescent iPSC reporter toolbox enables the visualization and isolation of key populations in forming kidney organoids, facilitating a range of applications, including cellular isolation, time-lapse imaging, protocol optimization, and lineage-tracing approaches. These tools offer promise for enhancing our understanding of this model system and its correspondence with human kidney morphogenesis. (Copyright © 2019 by the American Society of Nephrology.) |
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| معلومات مُعتمدة: | U01 DK107350 United States DK NIDDK NIH HHS; UH2 DK107344 United States DK NIDDK NIH HHS; UH3 DK107344 United States DK NIDDK NIH HHS |
| فهرسة مساهمة: | Keywords: kidney development; molecular biology; stem cell |
| تواريخ الأحداث: | Date Created: 20190908 Date Completed: 20200528 Latest Revision: 20201027 |
| رمز التحديث: | 20231215 |
| مُعرف محوري في PubMed: | PMC6779359 |
| DOI: | 10.1681/ASN.2019030303 |
| PMID: | 31492807 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1533-3450 |
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| DOI: | 10.1681/ASN.2019030303 |