Macrophage-like synoviocytes display phenotypic polymorphisms in a serum-free tissue-culture medium.
Synovial macrophages play an outstanding role in many rheumatic diseases. However, traditional serum-containing tissue-culture techniques hamper in vitro studies due to fibroblast activation not found in vivo. The objective of this study was to examine dissociated synovial cells in a macrophage-selective, serum-free tissue-culture medium. Osteoarthritis synovial tissue (n=11) was cultured in Iscove's Modified Dulbecco's Medium (IMDM) with 10% fetal bovine serum (FBS) and compared to a serum-free, insulin-supplemented medium. After 9-11 and 19-21 days in vitro, immunohistochemistry was performed for macrophage/lymphocyte markers and cell division. Cytokine profiles were determined by RT-PCR. In serum, cells with a bipolar morphology rapidly proliferated. Respectively, 14.34+/-12.94% and 13.25+/-12.66% expressed CD68 and HLA-DR. These markers further decreased after one passage. In serum-free medium, proliferation was infrequent, and cells with diverse morphologies expressed 83.10+/-6.80% and 55.03+/-6.88% CD68 and HLA-DR respectively. CD14 was rare, and lymphocytes were missing. Both cultures expressed interleukin-6 and interleukin-8. This novel serum-free method permits the culture of distinct CD68/HLA-DR associated phenotypes.
Seidel
Matthias F
MF
Department of Rheumatology, Medizinische Poliklinik der Universität Bonn, Wilhelmstrabe 35-37, Bonn, D-53111, Germany. Matthias.Seidel@ukb.uni-bonn.de
Koch
Franz-Walter
FW
Vetter
Hans
H
D000368:Aged; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D015415:Biomarkers / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D016895:Culture Media, Serum-Free / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D006684:HLA-DR Antigens / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008264:Macrophages / Q000145:classification* / Q000276:immunology; D008297:Male; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D013583:Synovial Membrane / Q000166:cytology*; D000097382:CD68 Molecule
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015415:Biomarkers; D016895:Culture Media, Serum-Free; D006684:HLA-DR Antigens; C067980:CD68 antigen, human; D000097382:CD68 Molecule