Scanu A, Oliviero F, Braghetto L, Ramonda R, Luisetto R, Calabrese F, Pozzuoli A, Punzi L
Cattedra e U.O.C. di Reumatologia, Università di Padova, Padova, Italia.
Reumatismo. 2007 Jan-Mar;59(1):66-70. doi: 10.4081/reumatismo.2007.66.
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).
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% foetal calf serum, 1% peni-streptomycin, 4 ng/ml of fibroblast grow factor and incubated at 37 degrees 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).
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.
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.
风湿性疾病发病机制的研究通常通过基于细胞培养模型的“体外”系统进行。获取新鲜人体组织的困难促使我们开发一种更简单的方法，从滑液（SF）中获取成纤维细胞样滑膜细胞。
从5例类风湿关节炎（RA）患者、4例骨关节炎（OA）患者和5例银屑病关节炎（PsA）患者的膝关节采集滑液。离心后获得的沉淀重悬于含有10%胎牛血清、1%青霉素-链霉素、4 ng/ml成纤维细胞生长因子的DMEM/HamF12中，并在T25培养瓶中于37℃孵育。滑膜细胞也通过外植体技术从新鲜滑膜（SM）中获得。两种类型的细胞均通过免疫细胞化学进行表征，并通过测量一氧化氮（NO）研究它们对合成尿酸钠晶体的炎症反应。
从2/5的RA滑液、4/4的OA滑液和5/5的PsA滑液培养物中获得了贴壁滑膜细胞。从滑液和滑膜中分离出的滑膜细胞均表达表面抗原CD90、CD55和细胞内脯氨酰-4-羟化酶。形态学上，细胞呈现典型的纺锤形成纤维细胞样外观。尿酸钠晶体在滑液滑膜细胞中诱导的NO水平与在滑膜滑膜细胞中获得的水平相似。
从滑液中获得的贴壁细胞表现出组织滑膜细胞的表型和反应性。由于滑液易于获取，当滑膜组织不能及时获得时，这种方法可能是一种有用的替代方法。
