Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis

Detalhes bibliográficos
Autor(a) principal: Zafari,Parisa
Data de Publicação: 2021
Outros Autores: Rafiei,Alireza, Faramarzi,Fatemeh, Ghaffari,Salman, Amiri,Aref Hosseinian, Taghadosi,Mahdi
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Revista da Associação Médica Brasileira (Online)
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-42302021001201654
Resumo: SUMMARY OBJECTIVE: Cell culture technology has become a popular method in the field of cell biology, pharmacology, and medical researches. Primary cells represent the normal physiological condition of human cells. Fibroblasts are the most common native cells of connective tissue that play a crucial role in the entire pathogenesis of various disorders, such as rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLSs), which overlie the loose connective tissue of the synovial sublining, are known to be the central mediators of joint damage. The most routine approach for the isolation of FLS is an enzymatic digestion of synovial tissue. This experimental study is designed to introduce an easy, fast, and high-throughput method compared with enzymatic digestion for isolation of FLS. METHODS: The synovial tissue and synovial fluid (SF) samples were collected from eight patients with RA who underwent routine knee replacement surgery. Synovial tissue was incubated with collagenase VIII enzyme, while SF was washed with a similar volume of phosphate-buffered saline. The cells were further subcultured and stored based on the standard protocols. The purity of isolated synoviocytes was confirmed using flow cytometry analysis. RESULTS: Isolation of FLS from SF was more successful with a faster rate, 3–5 days after culture. The morphological assessment and flow cytometry analysis confirmed the purity of SF-derived cells in passage 4. CONCLUSIONS: SF could be a more accessible source of FLS than synovial tissue. Obtaining primary FLS from SF is a simple, fast, and cost-effective way to have a large-scale cell during a short time.
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spelling Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritisCell cultureCell isolationFibroblast-like synoviocyteSynoviumSynovial fluidSUMMARY OBJECTIVE: Cell culture technology has become a popular method in the field of cell biology, pharmacology, and medical researches. Primary cells represent the normal physiological condition of human cells. Fibroblasts are the most common native cells of connective tissue that play a crucial role in the entire pathogenesis of various disorders, such as rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLSs), which overlie the loose connective tissue of the synovial sublining, are known to be the central mediators of joint damage. The most routine approach for the isolation of FLS is an enzymatic digestion of synovial tissue. This experimental study is designed to introduce an easy, fast, and high-throughput method compared with enzymatic digestion for isolation of FLS. METHODS: The synovial tissue and synovial fluid (SF) samples were collected from eight patients with RA who underwent routine knee replacement surgery. Synovial tissue was incubated with collagenase VIII enzyme, while SF was washed with a similar volume of phosphate-buffered saline. The cells were further subcultured and stored based on the standard protocols. The purity of isolated synoviocytes was confirmed using flow cytometry analysis. RESULTS: Isolation of FLS from SF was more successful with a faster rate, 3–5 days after culture. The morphological assessment and flow cytometry analysis confirmed the purity of SF-derived cells in passage 4. CONCLUSIONS: SF could be a more accessible source of FLS than synovial tissue. Obtaining primary FLS from SF is a simple, fast, and cost-effective way to have a large-scale cell during a short time.Associação Médica Brasileira2021-11-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-42302021001201654Revista da Associação Médica Brasileira v.67 n.11 2021reponame:Revista da Associação Médica Brasileira (Online)instname:Associação Médica Brasileira (AMB)instacron:AMB10.1590/1806-9282.20210706info:eu-repo/semantics/openAccessZafari,ParisaRafiei,AlirezaFaramarzi,FatemehGhaffari,SalmanAmiri,Aref HosseinianTaghadosi,Mahdieng2021-12-10T00:00:00Zoai:scielo:S0104-42302021001201654Revistahttps://ramb.amb.org.br/ultimas-edicoes/#https://old.scielo.br/oai/scielo-oai.php||ramb@amb.org.br1806-92820104-4230opendoar:2021-12-10T00:00Revista da Associação Médica Brasileira (Online) - Associação Médica Brasileira (AMB)false
dc.title.none.fl_str_mv Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
title Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
spellingShingle Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
Zafari,Parisa
Cell culture
Cell isolation
Fibroblast-like synoviocyte
Synovium
Synovial fluid
title_short Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
title_full Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
title_fullStr Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
title_full_unstemmed Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
title_sort Human fibroblast-like synoviocyte isolation matter: a comparison between cell isolation from synovial tissue and synovial fluid from patients with rheumatoid arthritis
author Zafari,Parisa
author_facet Zafari,Parisa
Rafiei,Alireza
Faramarzi,Fatemeh
Ghaffari,Salman
Amiri,Aref Hosseinian
Taghadosi,Mahdi
author_role author
author2 Rafiei,Alireza
Faramarzi,Fatemeh
Ghaffari,Salman
Amiri,Aref Hosseinian
Taghadosi,Mahdi
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Zafari,Parisa
Rafiei,Alireza
Faramarzi,Fatemeh
Ghaffari,Salman
Amiri,Aref Hosseinian
Taghadosi,Mahdi
dc.subject.por.fl_str_mv Cell culture
Cell isolation
Fibroblast-like synoviocyte
Synovium
Synovial fluid
topic Cell culture
Cell isolation
Fibroblast-like synoviocyte
Synovium
Synovial fluid
description SUMMARY OBJECTIVE: Cell culture technology has become a popular method in the field of cell biology, pharmacology, and medical researches. Primary cells represent the normal physiological condition of human cells. Fibroblasts are the most common native cells of connective tissue that play a crucial role in the entire pathogenesis of various disorders, such as rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLSs), which overlie the loose connective tissue of the synovial sublining, are known to be the central mediators of joint damage. The most routine approach for the isolation of FLS is an enzymatic digestion of synovial tissue. This experimental study is designed to introduce an easy, fast, and high-throughput method compared with enzymatic digestion for isolation of FLS. METHODS: The synovial tissue and synovial fluid (SF) samples were collected from eight patients with RA who underwent routine knee replacement surgery. Synovial tissue was incubated with collagenase VIII enzyme, while SF was washed with a similar volume of phosphate-buffered saline. The cells were further subcultured and stored based on the standard protocols. The purity of isolated synoviocytes was confirmed using flow cytometry analysis. RESULTS: Isolation of FLS from SF was more successful with a faster rate, 3–5 days after culture. The morphological assessment and flow cytometry analysis confirmed the purity of SF-derived cells in passage 4. CONCLUSIONS: SF could be a more accessible source of FLS than synovial tissue. Obtaining primary FLS from SF is a simple, fast, and cost-effective way to have a large-scale cell during a short time.
publishDate 2021
dc.date.none.fl_str_mv 2021-11-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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dc.language.iso.fl_str_mv eng
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dc.relation.none.fl_str_mv 10.1590/1806-9282.20210706
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dc.publisher.none.fl_str_mv Associação Médica Brasileira
publisher.none.fl_str_mv Associação Médica Brasileira
dc.source.none.fl_str_mv Revista da Associação Médica Brasileira v.67 n.11 2021
reponame:Revista da Associação Médica Brasileira (Online)
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reponame_str Revista da Associação Médica Brasileira (Online)
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repository.name.fl_str_mv Revista da Associação Médica Brasileira (Online) - Associação Médica Brasileira (AMB)
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