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The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time

BACKGROUND: Established therapies for managing kidney dysfunction such as kidney dialysis and transplantation are limited due to the shortage of compatible donated organs and high costs. Stem cell-based therapies are currently under investigation as an alternative treatment option. As amniotic fluid...

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Autores principales: Rahman, Md Shaifur, Spitzhorn, Lucas-Sebastian, Wruck, Wasco, Hagenbeck, Carsten, Balan, Percy, Graffmann, Nina, Bohndorf, Martina, Ncube, Audrey, Guillot, Pascale V., Fehm, Tanja, Adjaye, James
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5918774/
https://www.ncbi.nlm.nih.gov/pubmed/29695308
http://dx.doi.org/10.1186/s13287-018-0864-7
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author Rahman, Md Shaifur
Spitzhorn, Lucas-Sebastian
Wruck, Wasco
Hagenbeck, Carsten
Balan, Percy
Graffmann, Nina
Bohndorf, Martina
Ncube, Audrey
Guillot, Pascale V.
Fehm, Tanja
Adjaye, James
author_facet Rahman, Md Shaifur
Spitzhorn, Lucas-Sebastian
Wruck, Wasco
Hagenbeck, Carsten
Balan, Percy
Graffmann, Nina
Bohndorf, Martina
Ncube, Audrey
Guillot, Pascale V.
Fehm, Tanja
Adjaye, James
author_sort Rahman, Md Shaifur
collection PubMed
description BACKGROUND: Established therapies for managing kidney dysfunction such as kidney dialysis and transplantation are limited due to the shortage of compatible donated organs and high costs. Stem cell-based therapies are currently under investigation as an alternative treatment option. As amniotic fluid is composed of fetal urine harboring mesenchymal stem cells (AF-MSCs), we hypothesized that third-trimester amniotic fluid could be a novel source of renal progenitor and differentiated cells. METHODS: Human third-trimester amniotic fluid cells (AFCs) were isolated and cultured in distinct media. These cells were characterized as renal progenitor cells with respect to cell morphology, cell surface marker expression, transcriptome and differentiation into chondrocytes, osteoblasts and adipocytes. To test for renal function, a comparative albumin endocytosis assay was performed using AF-MSCs and commercially available renal cells derived from kidney biopsies. Comparative transcriptome analyses of first, second and third trimester-derived AF-MSCs were conducted to monitor expression of renal-related genes. RESULTS: Regardless of the media used, AFCs showed expression of pluripotency-associated markers such as SSEA4, TRA-1-60, TRA-1-81 and C-Kit. They also express the mesenchymal marker Vimentin. Immunophenotyping confirmed that third-trimester AFCs are bona fide MSCs. AF-MSCs expressed the master renal progenitor markers SIX2 and CITED1, in addition to typical renal proteins such as PODXL, LHX1, BRN1 and PAX8. Albumin endocytosis assays demonstrated the functionality of AF-MSCs as renal cells. Additionally, upregulated expression of BMP7 and downregulation of WT1, CD133, SIX2 and C-Kit were observed upon activation of WNT signaling by treatment with the GSK-3 inhibitor CHIR99201. Transcriptome analysis and semiquantitative PCR revealed increasing expression levels of renal-specific genes (e.g., SALL1, HNF4B, SIX2) with gestational time. Moreover, AF-MSCs shared more genes with human kidney cells than with native MSCs and gene ontology terms revealed involvement of biological processes associated with kidney morphogenesis. CONCLUSIONS: Third-trimester amniotic fluid contains AF-MSCs of renal origin and this novel source of kidney progenitors may have enormous future potentials for disease modeling, renal repair and drug screening. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0864-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-59187742018-04-30 The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time Rahman, Md Shaifur Spitzhorn, Lucas-Sebastian Wruck, Wasco Hagenbeck, Carsten Balan, Percy Graffmann, Nina Bohndorf, Martina Ncube, Audrey Guillot, Pascale V. Fehm, Tanja Adjaye, James Stem Cell Res Ther Research BACKGROUND: Established therapies for managing kidney dysfunction such as kidney dialysis and transplantation are limited due to the shortage of compatible donated organs and high costs. Stem cell-based therapies are currently under investigation as an alternative treatment option. As amniotic fluid is composed of fetal urine harboring mesenchymal stem cells (AF-MSCs), we hypothesized that third-trimester amniotic fluid could be a novel source of renal progenitor and differentiated cells. METHODS: Human third-trimester amniotic fluid cells (AFCs) were isolated and cultured in distinct media. These cells were characterized as renal progenitor cells with respect to cell morphology, cell surface marker expression, transcriptome and differentiation into chondrocytes, osteoblasts and adipocytes. To test for renal function, a comparative albumin endocytosis assay was performed using AF-MSCs and commercially available renal cells derived from kidney biopsies. Comparative transcriptome analyses of first, second and third trimester-derived AF-MSCs were conducted to monitor expression of renal-related genes. RESULTS: Regardless of the media used, AFCs showed expression of pluripotency-associated markers such as SSEA4, TRA-1-60, TRA-1-81 and C-Kit. They also express the mesenchymal marker Vimentin. Immunophenotyping confirmed that third-trimester AFCs are bona fide MSCs. AF-MSCs expressed the master renal progenitor markers SIX2 and CITED1, in addition to typical renal proteins such as PODXL, LHX1, BRN1 and PAX8. Albumin endocytosis assays demonstrated the functionality of AF-MSCs as renal cells. Additionally, upregulated expression of BMP7 and downregulation of WT1, CD133, SIX2 and C-Kit were observed upon activation of WNT signaling by treatment with the GSK-3 inhibitor CHIR99201. Transcriptome analysis and semiquantitative PCR revealed increasing expression levels of renal-specific genes (e.g., SALL1, HNF4B, SIX2) with gestational time. Moreover, AF-MSCs shared more genes with human kidney cells than with native MSCs and gene ontology terms revealed involvement of biological processes associated with kidney morphogenesis. CONCLUSIONS: Third-trimester amniotic fluid contains AF-MSCs of renal origin and this novel source of kidney progenitors may have enormous future potentials for disease modeling, renal repair and drug screening. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0864-7) contains supplementary material, which is available to authorized users. BioMed Central 2018-04-25 /pmc/articles/PMC5918774/ /pubmed/29695308 http://dx.doi.org/10.1186/s13287-018-0864-7 Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Rahman, Md Shaifur
Spitzhorn, Lucas-Sebastian
Wruck, Wasco
Hagenbeck, Carsten
Balan, Percy
Graffmann, Nina
Bohndorf, Martina
Ncube, Audrey
Guillot, Pascale V.
Fehm, Tanja
Adjaye, James
The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title_full The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title_fullStr The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title_full_unstemmed The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title_short The presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
title_sort presence of human mesenchymal stem cells of renal origin in amniotic fluid increases with gestational time
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5918774/
https://www.ncbi.nlm.nih.gov/pubmed/29695308
http://dx.doi.org/10.1186/s13287-018-0864-7
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