Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts
The composition of synovial fluid in rheumatoid arthritis (RA) is complex and strongly influences the microenvironment of joints and it is an inseparable element of the disease. Currently, “in vitro” studies are performed on RA cells cultured in the presence of either recombinant proinflammatory cyt...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2014-01-01
|
| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2014/702057 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832566877771005952 |
|---|---|
| author | Claudia Casnici Donatella Lattuada Noemi Tonna Katia Crotta Claudio Storini Fabio Bianco Marcello Claudio Truzzi Costantino Corradini Ornella Marelli |
| author_facet | Claudia Casnici Donatella Lattuada Noemi Tonna Katia Crotta Claudio Storini Fabio Bianco Marcello Claudio Truzzi Costantino Corradini Ornella Marelli |
| author_sort | Claudia Casnici |
| collection | DOAJ |
| description | The composition of synovial fluid in rheumatoid arthritis (RA) is complex and strongly influences the microenvironment of joints and it is an inseparable element of the disease. Currently, “in vitro” studies are performed on RA cells cultured in the presence of either recombinant proinflammatory cytokines-conditioned medium or medium alone. In this study, we evaluated the use of synovial fluid, derived from RA patients, as optimal culture condition to perform “in vitro” studies on RA synovial fibroblasts. We observed that synovial fluid is more effective in inducing cell proliferation with respect to TNF-alpha or culture medium alone. Spontaneous apoptosis in fibroblasts was also decreased in response to synovial fluid. The expression of proinflammatory cytokines in the presence of synovial fluid was significantly elevated with respect to cells cultured with TNF-alpha or medium, and the overall morphology of cells was also modified. In addition, modulation of intracellular calcium dynamics elicited in response to synovial fluid or TNF-alpha exposure is different and suggests a role for the purinergic signalling in the modulation of the effects. These results emphasize the importance of using RA synovial fluid in “in vitro” studies involving RA cells, in order to reproduce faithfully the physiopathological environmental characteristic of RA joints. |
| format | Article |
| id | doaj-art-0f0981c73fcc41ee8b10025271247ca6 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-0f0981c73fcc41ee8b10025271247ca62025-02-03T01:02:55ZengWileyMediators of Inflammation0962-93511466-18612014-01-01201410.1155/2014/702057702057Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial FibroblastsClaudia Casnici0Donatella Lattuada1Noemi Tonna2Katia Crotta3Claudio Storini4Fabio Bianco5Marcello Claudio Truzzi6Costantino Corradini7Ornella Marelli8Department of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, ItalyDepartment of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, ItalyNeuro-Zone s.r.l, Viale Ortles 22/4, 20139 Milan, ItalyDepartment of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, ItalyOrthopaedic Hospital Gaetano Pini, Piazza A. Ferrari 1, 20122 Milan, ItalyNeuro-Zone s.r.l, Viale Ortles 22/4, 20139 Milan, ItalySanipedia s.r.l, Via Podgora 7, 20122 Milan, ItalySanipedia s.r.l, Via Podgora 7, 20122 Milan, ItalyDepartment of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, ItalyThe composition of synovial fluid in rheumatoid arthritis (RA) is complex and strongly influences the microenvironment of joints and it is an inseparable element of the disease. Currently, “in vitro” studies are performed on RA cells cultured in the presence of either recombinant proinflammatory cytokines-conditioned medium or medium alone. In this study, we evaluated the use of synovial fluid, derived from RA patients, as optimal culture condition to perform “in vitro” studies on RA synovial fibroblasts. We observed that synovial fluid is more effective in inducing cell proliferation with respect to TNF-alpha or culture medium alone. Spontaneous apoptosis in fibroblasts was also decreased in response to synovial fluid. The expression of proinflammatory cytokines in the presence of synovial fluid was significantly elevated with respect to cells cultured with TNF-alpha or medium, and the overall morphology of cells was also modified. In addition, modulation of intracellular calcium dynamics elicited in response to synovial fluid or TNF-alpha exposure is different and suggests a role for the purinergic signalling in the modulation of the effects. These results emphasize the importance of using RA synovial fluid in “in vitro” studies involving RA cells, in order to reproduce faithfully the physiopathological environmental characteristic of RA joints.http://dx.doi.org/10.1155/2014/702057 |
| spellingShingle | Claudia Casnici Donatella Lattuada Noemi Tonna Katia Crotta Claudio Storini Fabio Bianco Marcello Claudio Truzzi Costantino Corradini Ornella Marelli Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts Mediators of Inflammation |
| title | Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts |
| title_full | Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts |
| title_fullStr | Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts |
| title_full_unstemmed | Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts |
| title_short | Optimized “In Vitro” Culture Conditions for Human Rheumatoid Arthritis Synovial Fibroblasts |
| title_sort | optimized in vitro culture conditions for human rheumatoid arthritis synovial fibroblasts |
| url | http://dx.doi.org/10.1155/2014/702057 |
| work_keys_str_mv | AT claudiacasnici optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT donatellalattuada optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT noemitonna optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT katiacrotta optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT claudiostorini optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT fabiobianco optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT marcelloclaudiotruzzi optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT costantinocorradini optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts AT ornellamarelli optimizedinvitrocultureconditionsforhumanrheumatoidarthritissynovialfibroblasts |