Cargando…

Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment

BACKGROUND: Bone marrow and adipose tissues are known sources of mesenchymal stem cells (MSCs) in horses; however, synovial tissues might be a promising alternative. The aim of this study was to evaluate phenotypic characteristics and differentiation potential of equine MSCs from synovial fluid (SF)...

Descripción completa

Detalles Bibliográficos
Autores principales: Fülber, Joice, Maria, Durvanei A., Silva, Luis Cláudio Lopes Correia da, Massoco, Cristina O., Agreste, Fernanda, Baccarin, Raquel Y. Arantes
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4779201/
https://www.ncbi.nlm.nih.gov/pubmed/26944403
http://dx.doi.org/10.1186/s13287-016-0294-3
_version_ 1782419593630842880
author Fülber, Joice
Maria, Durvanei A.
Silva, Luis Cláudio Lopes Correia da
Massoco, Cristina O.
Agreste, Fernanda
Baccarin, Raquel Y. Arantes
author_facet Fülber, Joice
Maria, Durvanei A.
Silva, Luis Cláudio Lopes Correia da
Massoco, Cristina O.
Agreste, Fernanda
Baccarin, Raquel Y. Arantes
author_sort Fülber, Joice
collection PubMed
description BACKGROUND: Bone marrow and adipose tissues are known sources of mesenchymal stem cells (MSCs) in horses; however, synovial tissues might be a promising alternative. The aim of this study was to evaluate phenotypic characteristics and differentiation potential of equine MSCs from synovial fluid (SF) and synovial membrane (SM) of healthy joints (SF-H and SM-H), joints with osteoarthritis (SF-OA and SM-OA) and joints with osteochondritis dissecans (SF-OCD and SM-OCD) to determine the most suitable synovial source for an allogeneic therapy cell bank. METHODS: Expression of the markers CD90, CD105, CD44, and CD34 in SF-H, SM-H, SF-OA, SM-OA, SF-OCD and SM-OCD was verified by flow cytometry, and expression of cytokeratin, vimentin, PGP 9.5, PCNA, lysozyme, nanog, and Oct4 was verified by immunocytochemistry. MSCs were cultured and evaluated for their chondrogenic, osteogenic and adipogenic differentiation potential. Final quantification of extracellular matrix and mineralized matrix was determined using AxioVision software. A tumorigenicity test was conducted in Balb-C(nu/nu) mice to verify the safety of the MSCs from these sources. RESULTS: Cultured cells from SF and SM exhibited fibroblastoid morphology and the ability to adhere to plastic. The time elapsed between primary culture and the third passage was approximately 73 days for SF-H, 89 days for SF-OCD, 60 days for SF-OA, 68 days for SM-H, 57 days for SM-OCD and 54 days for SM-OA. The doubling time for SF-OCD was higher than that for other cells at the first passage (P < 0.05). MSCs from synovial tissues showed positive expression of the markers CD90, CD44, lysozyme, PGP 9.5, PCNA and vimentin and were able to differentiate into chondrogenic (21 days) and osteogenic (21 days) lineages, and, although poorly, into adipogenic lineages (14 days). The areas staining positive for extracellular matrix in the SF-H and SM-H groups were larger than those in the SF-OA and SM-OA groups (P < 0.05). The positive mineralized matrix area in the SF-H group was larger than those in all the other groups (P < 0.05). The studied cells exhibited no tumorigenic effects. CONCLUSIONS: SF and SM are viable sources of equine MSCs. All sources studied provide suitable MSCs for an allogeneic therapy cell bank; nevertheless, MSCs from healthy joints may be preferable for cell banking purposes because they exhibit better chondrogenic differentiation capacity.
format Online
Article
Text
id pubmed-4779201
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-47792012016-03-06 Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment Fülber, Joice Maria, Durvanei A. Silva, Luis Cláudio Lopes Correia da Massoco, Cristina O. Agreste, Fernanda Baccarin, Raquel Y. Arantes Stem Cell Res Ther Research BACKGROUND: Bone marrow and adipose tissues are known sources of mesenchymal stem cells (MSCs) in horses; however, synovial tissues might be a promising alternative. The aim of this study was to evaluate phenotypic characteristics and differentiation potential of equine MSCs from synovial fluid (SF) and synovial membrane (SM) of healthy joints (SF-H and SM-H), joints with osteoarthritis (SF-OA and SM-OA) and joints with osteochondritis dissecans (SF-OCD and SM-OCD) to determine the most suitable synovial source for an allogeneic therapy cell bank. METHODS: Expression of the markers CD90, CD105, CD44, and CD34 in SF-H, SM-H, SF-OA, SM-OA, SF-OCD and SM-OCD was verified by flow cytometry, and expression of cytokeratin, vimentin, PGP 9.5, PCNA, lysozyme, nanog, and Oct4 was verified by immunocytochemistry. MSCs were cultured and evaluated for their chondrogenic, osteogenic and adipogenic differentiation potential. Final quantification of extracellular matrix and mineralized matrix was determined using AxioVision software. A tumorigenicity test was conducted in Balb-C(nu/nu) mice to verify the safety of the MSCs from these sources. RESULTS: Cultured cells from SF and SM exhibited fibroblastoid morphology and the ability to adhere to plastic. The time elapsed between primary culture and the third passage was approximately 73 days for SF-H, 89 days for SF-OCD, 60 days for SF-OA, 68 days for SM-H, 57 days for SM-OCD and 54 days for SM-OA. The doubling time for SF-OCD was higher than that for other cells at the first passage (P < 0.05). MSCs from synovial tissues showed positive expression of the markers CD90, CD44, lysozyme, PGP 9.5, PCNA and vimentin and were able to differentiate into chondrogenic (21 days) and osteogenic (21 days) lineages, and, although poorly, into adipogenic lineages (14 days). The areas staining positive for extracellular matrix in the SF-H and SM-H groups were larger than those in the SF-OA and SM-OA groups (P < 0.05). The positive mineralized matrix area in the SF-H group was larger than those in all the other groups (P < 0.05). The studied cells exhibited no tumorigenic effects. CONCLUSIONS: SF and SM are viable sources of equine MSCs. All sources studied provide suitable MSCs for an allogeneic therapy cell bank; nevertheless, MSCs from healthy joints may be preferable for cell banking purposes because they exhibit better chondrogenic differentiation capacity. BioMed Central 2016-03-05 /pmc/articles/PMC4779201/ /pubmed/26944403 http://dx.doi.org/10.1186/s13287-016-0294-3 Text en © Fülber et al. 2016 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
Fülber, Joice
Maria, Durvanei A.
Silva, Luis Cláudio Lopes Correia da
Massoco, Cristina O.
Agreste, Fernanda
Baccarin, Raquel Y. Arantes
Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title_full Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title_fullStr Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title_full_unstemmed Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title_short Comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
title_sort comparative study of equine mesenchymal stem cells from healthy and injured synovial tissues: an in vitro assessment
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4779201/
https://www.ncbi.nlm.nih.gov/pubmed/26944403
http://dx.doi.org/10.1186/s13287-016-0294-3
work_keys_str_mv AT fulberjoice comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment
AT mariadurvaneia comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment
AT silvaluisclaudiolopescorreiada comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment
AT massococristinao comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment
AT agrestefernanda comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment
AT baccarinraquelyarantes comparativestudyofequinemesenchymalstemcellsfromhealthyandinjuredsynovialtissuesaninvitroassessment