Successful in vitro culture of synovial explants from rheumatoid arthritis at the air-liquid interface.
Successful in vitro culture of synovial explants from rheumatoid arthritis at the air-liquid interface.
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在气液界面成功体外培养类风湿性关节炎滑膜外植体。
DOI：
10.1002/art.39019
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Yamada H.
中科院分区：
文献类型：
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作者：
Sakuraba K;Fujimura K;Nakashima Y;Okazaki K;Fukushi JI;Ohishi M;Oyamada A;Esaki Y;Miyahara H;Iwamoto Y;Yoshikai Y;Yamada H.
目的建立类风湿性关节炎（RA）患者滑膜组织外植体的培养方法将10例RA患者的滑膜组织外植体进行气液界面培养或浸没在培养基中培养。作为对照，将滑膜外植体皮下移植到SCID小鼠中。在不同时间点收获滑膜外植体，并进行组织学或流式细胞术分析。通过酶联免疫吸附试验测定培养上清液中的细胞因子水平。英夫利西单抗被添加到空气-液体界面culture.ResultsThe RA滑膜炎的组织学特征，包括一个增生的衬里层和存在的细胞浸润的亚衬里层，保持在空气-液体界面培养的滑膜组织外植体肿瘤坏死因子α封锁的影响。在从SCID-HuRAg小鼠收获的滑膜移植物中，细胞浸润在下层中保持良好，但衬里层丢失。在气液界面培养后，活的CD 4 + T细胞和巨噬细胞丰富，但在浸没培养后几乎不存在。此外，空气-液体界面培养中的滑膜组织外植体产生白细胞介素-6（IL-6）和IL-8的时间较长。英夫利西单抗的加入有效地减少了细胞因子production.ConclusionRA滑膜外植体可以维持数周使用气液界面培养。这种简单的培养系统可能有助于分析RA滑膜炎的发病机制和开发抗风湿药物。
ObjectiveTo establish a method to culture synovial tissue explants from patients with rheumatoid arthritis (RA).MethodsSynovial tissue explants obtained from 10 patients with RA were cultured at the air–liquid interface or were submerged in culture medium. As a control, synovial explants were engrafted subcutaneously into SCID mice. The synovial explants were harvested at different time points, and histologic or flow cytometric analysis was performed. Cytokine levels in the culture supernatants were measured by enzyme‐linked immunosorbent assay. Infliximab was added to the air–liquid interface culture to evaluate the effect of tumor necrosis factor α blockade on inflammatory cytokine production.ResultsThe histologic features of RA synovitis, including a hyperplastic lining layer and the presence of cellular infiltrate in the sublining layer, were maintained in synovial tissue explants in air–liquid interface culture. In synovial grafts harvested from SCID‐HuRAg mice, the cellular infiltrate was well maintained in the sublining, but the lining layer was lost. Viable CD4+ T cells and macrophages were abundant after air–liquid interface culture but were virtually absent after submerged culture. Furthermore, synovial tissue explants in air–liquid interface culture produced interleukin‐6 (IL‐6) and IL‐8 for a prolonged period of time. The addition of infliximab effectively reduced cytokine production.ConclusionRA synovial explants can be maintained for weeks using an air–liquid interface culture. This simple culture system might be useful for analyzing the pathogenesis of RA synovitis and for developing antirheumatic drugs.