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Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes

OBJECTIVES: Articular cartilage is vulnerable to injuries and undergoes an irreversible degenerative process. The use of amniotic fluid mesenchymal stromal stem cells for the reconstruction of articular cartilage is a promising therapeutic alternative. The aim of this study was to investigate the ch...

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Autores principales: Zuliani, Carolina Coli, Bombini, Mariana Freschi, de Andrade, Kleber Cursino, Mamoni, Ronei, Pereira, Ana Helena, Coimbra, Ibsen Bellini
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5866404/
https://www.ncbi.nlm.nih.gov/pubmed/29641802
http://dx.doi.org/10.6061/clinics/2018/e268
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author Zuliani, Carolina Coli
Bombini, Mariana Freschi
de Andrade, Kleber Cursino
Mamoni, Ronei
Pereira, Ana Helena
Coimbra, Ibsen Bellini
author_facet Zuliani, Carolina Coli
Bombini, Mariana Freschi
de Andrade, Kleber Cursino
Mamoni, Ronei
Pereira, Ana Helena
Coimbra, Ibsen Bellini
author_sort Zuliani, Carolina Coli
collection PubMed
description OBJECTIVES: Articular cartilage is vulnerable to injuries and undergoes an irreversible degenerative process. The use of amniotic fluid mesenchymal stromal stem cells for the reconstruction of articular cartilage is a promising therapeutic alternative. The aim of this study was to investigate the chondrogenic potential of amniotic fluid mesenchymal stromal stem cells from human amniotic fluid from second trimester pregnant women in a micromass system (high-density cell culture) with TGF-β3 for 21 days. METHODS: Micromass was performed using amniotic fluid mesenchymal stromal stem cells previously cultured in a monolayer. Chondrocytes from adult human normal cartilage were used as controls. After 21 days, chondrogenic potential was determined by measuring the expression of genes, such as SOX-9, type II collagen and aggrecan, in newly differentiated cells by real-time PCR (qRT-PCR). The production of type II collagen protein was observed by western blotting. Immunohistochemistry analysis was also performed to detect collagen type II and aggrecan. This study was approved by the local ethics committee. RESULTS: SOX-9, aggrecan and type II collagen were expressed in newly differentiated chondrocytes. The expression of SOX-9 was significantly higher in newly differentiated chondrocytes than in adult cartilage. Collagen type II protein was also detected. CONCLUSION: We demonstrate that stem cells from human amniotic fluid are a suitable source for chondrogenesis when cultured in a micromass system. amniotic fluid mesenchymal stromal stem cells are an extremely viable source for clinical applications, and our results suggest the possibility of using human amniotic fluid as a source of mesenchymal stem cells.
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spelling pubmed-58664042018-03-25 Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes Zuliani, Carolina Coli Bombini, Mariana Freschi de Andrade, Kleber Cursino Mamoni, Ronei Pereira, Ana Helena Coimbra, Ibsen Bellini Clinics (Sao Paulo) Original Article OBJECTIVES: Articular cartilage is vulnerable to injuries and undergoes an irreversible degenerative process. The use of amniotic fluid mesenchymal stromal stem cells for the reconstruction of articular cartilage is a promising therapeutic alternative. The aim of this study was to investigate the chondrogenic potential of amniotic fluid mesenchymal stromal stem cells from human amniotic fluid from second trimester pregnant women in a micromass system (high-density cell culture) with TGF-β3 for 21 days. METHODS: Micromass was performed using amniotic fluid mesenchymal stromal stem cells previously cultured in a monolayer. Chondrocytes from adult human normal cartilage were used as controls. After 21 days, chondrogenic potential was determined by measuring the expression of genes, such as SOX-9, type II collagen and aggrecan, in newly differentiated cells by real-time PCR (qRT-PCR). The production of type II collagen protein was observed by western blotting. Immunohistochemistry analysis was also performed to detect collagen type II and aggrecan. This study was approved by the local ethics committee. RESULTS: SOX-9, aggrecan and type II collagen were expressed in newly differentiated chondrocytes. The expression of SOX-9 was significantly higher in newly differentiated chondrocytes than in adult cartilage. Collagen type II protein was also detected. CONCLUSION: We demonstrate that stem cells from human amniotic fluid are a suitable source for chondrogenesis when cultured in a micromass system. amniotic fluid mesenchymal stromal stem cells are an extremely viable source for clinical applications, and our results suggest the possibility of using human amniotic fluid as a source of mesenchymal stem cells. Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo 2018-03-24 2018 /pmc/articles/PMC5866404/ /pubmed/29641802 http://dx.doi.org/10.6061/clinics/2018/e268 Text en Copyright © 2018 CLINICS http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium or format, provided the original work is properly cited.
spellingShingle Original Article
Zuliani, Carolina Coli
Bombini, Mariana Freschi
de Andrade, Kleber Cursino
Mamoni, Ronei
Pereira, Ana Helena
Coimbra, Ibsen Bellini
Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title_full Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title_fullStr Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title_full_unstemmed Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title_short Micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
title_sort micromass cultures are effective for differentiation of human amniotic fluid stem cells into chondrocytes
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5866404/
https://www.ncbi.nlm.nih.gov/pubmed/29641802
http://dx.doi.org/10.6061/clinics/2018/e268
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