Reconstruction culture system simulating human synovium: a three-dimensional collagen gel culture of synoviocytes.
We developed a reconstruction culture system simulating human synovium to investigate the synoviocytes in a more physiological condition. This system with two types of dishes provided the two separated spaces corresponding to the joint cavity and nutrient vessels. The isolated normal synoviocytes were cultured in type-I collagen gel with a layer-seeded on top in a smaller inner dish with a porous bottom, which was placed in a larger outer dish. We added hyaluronic acid solution to the space on the gel to make an environment close to the physiological joint cavity. The space between the inner and outer dishes was filled with a complete medium to nourish the synoviocytes through the porous filter (Experiment 1). In addition, to examine cell-to-cell interaction, we created a co-culture model by mixing synoviocytes and peripheral blood mononuclear cells in the collagen gel matrix (Experiment 2). In this way we could reconstruct the synovium in vitro. The synoviocytes could survive and maintain their characteristics for four weeks of culture. In Experiment 1, almost all the cells were similar to type B synovial cells by histological, immunohistochemical and electron microscopic observations. In Experiment 2, the spherical cells in abundance of lysosomes like type A synovial cells appeared sporadically in the lining layer. Immunohistochemically, the majority of cultured cells expressed CD68 and matrix metalloproteinase 3. This culture system promises to be of use in investigating the pathogenesis of various joint diseases.
Taguchi
K
K
Department of Orthopaedic Surgery, Nagasaki University School of Medicine, Japan.
Hirano
T
T
Iwasaki
K
K
Sugihara
H
H
D000293:Adolescent; D000328:Adult; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D018920:Coculture Techniques; D003094:Collagen / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D005782:Gels; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology; D013583:Synovial Membrane / Q000166:cytology* / Q000648:ultrastructure
D003470:Culture Media; D005782:Gels; D003094:Collagen