Synoviocyte cultures from synovial fluid

Abstract

The study of the pathogenetic mechanisms of rheumatic diseases is in general carried out through “in vitro” systems based on cellular cultures models. The difficulties to achieve fresh human tissue prompted us to develop a simpler method to obtain fibroblast-like synovial cells from synovial fluid (SF). Methods: SF was collected from the knees of 5 patients with rheumatoid arthritis (RA), 4 with osteoarthritis (OA) and 5 with psoriatic arthritis (PsA). The pellet obtained after centrifugation was resuspended in DMEM/HamF12 containing 10% fetal calf serum, 1% peni-streptomicin, 4ng/ml of fibroblast grow factor and incubated at 37°C in T25 culture flasks. Synoviocytes were also obtained from fresh synovial membranes (SM) by explants technique. Both types of cells were characterized by immunocytochemistry and their inflammatory response to synthetic monosodium urate crystals was studied through the measurement of nitric oxide (NO). Results: Adherent synoviocytes were obtained from the culture of 2/5 SF from RA, 4/4 SF from OA and 5/5 SF from PsA. Synoviocytes isolated from both SF and SM expressed surface antigens CD90, CD55, and the intracellular prolyl- 4-hydroxylase. Morphologically, the cells showed the typical spindle-shape fibroblast-like appearance. NO levels induced by UMS crystals in SF synoviocytes were similar to those obtained in SM synoviocytes. Conclusion: Adherent cells obtained from SF showed the phenotype and the reactivity of tissue synoviocytes. Due to the easy accessibility of SF, this method may represents an useful alternative when synovial tissues is not promptly available

Interleukin 37 limits monosodium urate crystal-induced innate immune responses in human and murine models of gout

“…Synovial cells (containing macrophage-like and fibroblast-like synoviocytes) were isolated from tissue explants and cultured as previously described [10]. Briefly, tissues were minced and incubated with a solution containing 0.15 mg/ml DNase, 0.15 mg/ml hyaluronidase (type I-S), and 1 mg/ml collagenase (type IA) (Sigma-Aldrich) in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen, Carlsbad, CA, USA) for 1 h at 37 °C.…”

Molecular basis of oxidative stress in gouty arthropathy

“…It is possible that the high production of antioxidants enzymes are due to the Nrf2 activation by ROS, like in the TPH-1 cells activation exposed to MSU with the intention of compensating the cellular stress state [34]. In the particular case of gout and other microcrystalline arthropathies, it is known that crystals increase the production of NO − [35], and if it combines with O 2 − , peroxynitrite (ONOO − ) is produced, affecting cellular proliferation, degradation of connective tissue structures, and degeneration of joint tissue [36][37][38].…”

Altered expression of TPP1 in fibroblast-like synovial cells might be involved in the pathogenesis of rheumatoid arthritis

“…Lines inside the boxes represent the median. Statistical signiWcance was set at P · 0.05 Studies have demonstrated that FLS obtained from synovial Xuid (SF) showed the phenotype and the reactivity of synovial tissue FLS [28,33]. Neidhart et al [53] demonstrated that similar to tissue-derived RA FLS, FLS from RA synovial Xuid, which contain "Xoating" anchorage-independent Wbroblast-like cells, mediate cartilage destruction independent of the hyperplastic synovial tissue.…”

“…Adherent cells were fed with the same media every 3-5 days and passaged every 7-14 days. The methods were described previously by Scanu et al [28]. Normal synovial tissues were obtained from 13 patients undergoing joint surgery after injury.…”

“…The round-shaped cells were more resistant to trypsin digestion than the spindle-shaped cells and remained in the Xask. After 2-3 passages, 90% of adherent cells are spindle-shaped cells, that is Wbroblast-like synovial cells (FLS), as described in previous studies [28,33]. FLS were used from 3 to 5 passages in the subsequent experiments.…”