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

Clinics. 2018;73(0) DOI 10.6061/clinics/2018/e268

 

Journal Homepage

Journal Title: Clinics

ISSN: 1807-5932 (Print); 1980-5322 (Online)

Publisher: Faculdade de Medicina / USP

LCC Subject Category: Medicine: Medicine (General)

Country of publisher: Brazil

Language of fulltext: English

Full-text formats available: PDF, HTML, XML

 

AUTHORS


Carolina Coli Zuliani

Mariana Freschi Bombini

Kleber Cursino de Andrade

Ronei Mamoni

Ana Helena Pereira

Ibsen Bellini Coimbra

EDITORIAL INFORMATION

Peer review

Editorial Board

Instructions for authors

Time From Submission to Publication: 7 weeks

 

Abstract | Full Text

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.