Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level
During nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a combination of surface markers. However, upon cultur...
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doaj-d8cb8a41442a4e62802a6d3cc08bb5552020-11-24T21:11:23ZengElsevierStem Cell Reports2213-67112017-07-019127929110.1016/j.stemcr.2017.04.026Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell LevelNaomi Pode-Shakked0Rotem Gershon1Gal Tam2Dorit Omer3Yehudit Gnatek4Itamar Kanter5Sarit Oriel6Guy Katz7Orit Harari-Steinberg8Tomer Kalisky9Benjamin Dekel10Pediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelFaculty of Engineering and Bar-Ilan Institute of Nanotechnology and Advanced Materials (BINA), Bar-Ilan University, Ramat Gan 5290002, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelFaculty of Engineering and Bar-Ilan Institute of Nanotechnology and Advanced Materials (BINA), Bar-Ilan University, Ramat Gan 5290002, IsraelFaculty of Engineering and Bar-Ilan Institute of Nanotechnology and Advanced Materials (BINA), Bar-Ilan University, Ramat Gan 5290002, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelFaculty of Engineering and Bar-Ilan Institute of Nanotechnology and Advanced Materials (BINA), Bar-Ilan University, Ramat Gan 5290002, IsraelPediatric Stem Cell Research Institute, Edmond and Lily Safra Children's Hospital, Sheba Medical Center, Tel-Hashomer 52621, IsraelDuring nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a combination of surface markers. However, upon culture nephron progenitors differentiated and could not be robustly maintained in vitro. Here, by culturing hFK in a modified medium used for in vitro growth of mouse nephron progenitors, and by dissection of NCAM+/CD133− progenitor cells according to EpCAM expression (NCAM+/CD133−/EpCAM−, NCAM+/CD133−/EpCAMdim, NCAM+/CD133−/EpCAMbright), we show at single-cell resolution a preservation of uninduced and induced cap mesenchyme as well as a transitioning mesenchymal-epithelial state. Concomitantly, differentiating and differentiated epithelial lineages are also maintained. In vitro expansion of discrete stages of early human nephrogenesis in nephron stem cell cultures may be used for drug screening on a full repertoire of developing kidney cells and for prospective isolation of mesenchymal or epithelial renal lineages for regenerative medicine.http://www.sciencedirect.com/science/article/pii/S2213671117301807stem cellskidney stem/progenitor cellsrenal developmentstem cell markersWilms' tumorsingle cell gene expression analysiscancer stem cells |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Naomi Pode-Shakked Rotem Gershon Gal Tam Dorit Omer Yehudit Gnatek Itamar Kanter Sarit Oriel Guy Katz Orit Harari-Steinberg Tomer Kalisky Benjamin Dekel |
spellingShingle |
Naomi Pode-Shakked Rotem Gershon Gal Tam Dorit Omer Yehudit Gnatek Itamar Kanter Sarit Oriel Guy Katz Orit Harari-Steinberg Tomer Kalisky Benjamin Dekel Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level Stem Cell Reports stem cells kidney stem/progenitor cells renal development stem cell markers Wilms' tumor single cell gene expression analysis cancer stem cells |
author_facet |
Naomi Pode-Shakked Rotem Gershon Gal Tam Dorit Omer Yehudit Gnatek Itamar Kanter Sarit Oriel Guy Katz Orit Harari-Steinberg Tomer Kalisky Benjamin Dekel |
author_sort |
Naomi Pode-Shakked |
title |
Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level |
title_short |
Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level |
title_full |
Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level |
title_fullStr |
Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level |
title_full_unstemmed |
Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level |
title_sort |
evidence of in vitro preservation of human nephrogenesis at the single-cell level |
publisher |
Elsevier |
series |
Stem Cell Reports |
issn |
2213-6711 |
publishDate |
2017-07-01 |
description |
During nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a combination of surface markers. However, upon culture nephron progenitors differentiated and could not be robustly maintained in vitro. Here, by culturing hFK in a modified medium used for in vitro growth of mouse nephron progenitors, and by dissection of NCAM+/CD133− progenitor cells according to EpCAM expression (NCAM+/CD133−/EpCAM−, NCAM+/CD133−/EpCAMdim, NCAM+/CD133−/EpCAMbright), we show at single-cell resolution a preservation of uninduced and induced cap mesenchyme as well as a transitioning mesenchymal-epithelial state. Concomitantly, differentiating and differentiated epithelial lineages are also maintained. In vitro expansion of discrete stages of early human nephrogenesis in nephron stem cell cultures may be used for drug screening on a full repertoire of developing kidney cells and for prospective isolation of mesenchymal or epithelial renal lineages for regenerative medicine. |
topic |
stem cells kidney stem/progenitor cells renal development stem cell markers Wilms' tumor single cell gene expression analysis cancer stem cells |
url |
http://www.sciencedirect.com/science/article/pii/S2213671117301807 |
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