title: Optimized "in vitro" culture conditions for human rheumatoid arthritis synovial fibroblasts.
abstract: 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.
authors.affiliation: Department of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, Italy ; Neuro-Zone s.r.l, Viale Ortles 22/4, 20139 Milan, Italy.
authors.forename: Claudia
authors.identifier: 0000-0002-5478-0239
authors.initials: C
authors.lastname: Casnici
authors.affiliation: Department of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, Italy.
authors.forename: Donatella
authors.initials: D
authors.lastname: Lattuada
authors.affiliation: Neuro-Zone s.r.l, Viale Ortles 22/4, 20139 Milan, Italy.
authors.forename: Noemi
authors.identifier: 0000-0002-4788-2413
authors.initials: N
authors.lastname: Tonna
authors.affiliation: Department of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, Italy.
authors.forename: Katia
authors.initials: K
authors.lastname: Crotta
authors.affiliation: Orthopaedic Hospital Gaetano Pini, Piazza A. Ferrari 1, 20122 Milan, Italy.
authors.forename: Claudio
authors.identifier: 0000-0002-5185-6277
authors.initials: C
authors.lastname: Storini
authors.affiliation: Neuro-Zone s.r.l, Viale Ortles 22/4, 20139 Milan, Italy ; Orthopaedic Hospital Gaetano Pini, Piazza A. Ferrari 1, 20122 Milan, Italy.
authors.forename: Fabio
authors.identifier: 0000-0001-9058-756X
authors.initials: F
authors.lastname: Bianco
authors.affiliation: Sanipedia s.r.l, Via Podgora 7, 20122 Milan, Italy.
authors.forename: Marcello Claudio
authors.initials: MC
authors.lastname: Truzzi
authors.affiliation: Sanipedia s.r.l, Via Podgora 7, 20122 Milan, Italy ; Centro Studi e Ricerche in Traumatologia dello Sport, University of Milan c/o I Divisione, Istituto Ortopedico G. Pini, Piazza A. Ferrari 1, 20122 Milan, Italy.
authors.forename: Costantino
authors.initials: C
authors.lastname: Corradini
authors.affiliation: Department of Medical Biotechnologies and Translational Medicine (BIOMETRA), University of Milan, Via Vanvitelli 32, 20129 Milan, Italy.
authors.forename: Ornella
authors.initials: O
authors.lastname: Marelli
mesh_terms: D000199:Actins / Q000378:metabolism; D017209:Apoptosis; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D049109:Cell Proliferation; D003470:Culture Media / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D007249:Inflammation; D013582:Synovial Fluid / Q000378:metabolism; D013583:Synovial Membrane / Q000737:chemistry* / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000199:Actins; D003470:Culture Media; D016207:Cytokines; D014409:Tumor Necrosis Factor-alpha; D002118:Calcium