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Báo cáo y học: "Flow cytometric characterization of freshly isolated and culture expanded human synovial cell populations in patients with chronic arthritis"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học General Psychiatry cung cấp cho các bạn kiến thức về ngành y đề tài: Flow cytometric characterization of freshly isolated and culture expanded human synovial cell populations in patients with chronic arthritis. | Van Landuyt et al. Arthritis Research Therapy 2010 12 R15 http arthritis-research.eom content 12 1 R15 RESEARCH ARTICLE Open Access Flow cytometric characterization of freshly isolated and culture expanded human synovial cell populations in patients with chronic arthritis Kristel B Van Landuyt1 Elena A Jones2 Dennis McGonagle2 Frank P Luyten1 Rik J Lories1 Abstract Introduction The synovium is a major target tissue in chronic arthritis and is intensively studied at the cellular and molecular level. The aim of this study was to develop flow cytometry for the quantitative analysis of synovial cell populations pre and post culture and to characterize mesenchymal cell populations residing in the inflammatory synovium. Methods Knee synovium biopsies from 39 patients with chronic arthritis and from 15 controls were treated in a short standardized tissue digestion procedure. Stored thawed digests were routinely analyzed with flow cytometry including live-dead staining and use of the markers CD45 CD3 CD14 CD20 CD34 CD73 CD105 CD90 CD146 CD163 and HLA-DR to distinguish inflammatory and stromal cells. The influence of the digestion method on the detection of the different surface markers was studied separately. In addition we studied the presence of a specific cell population hypothesized to be mesenchymal stem cells MSC based on the CD271 marker. Cell expansion cultures were set up and a MSC-related surface marker profile in passages 3 and 6 was obtained. Immunohistochemistry for CD34 and von Willebrand factor vWF was done to obtain additional data on synovium vascularity. Results The cell yield and viability normalized to tissue weight were significantly higher in inflammatory arthritis than in controls. Within the hematopoietic CD45-positive populations we found no differences in relative amounts of macrophages T-lymphocytes and B-lymphocytes between patient groups. Within the CD45-negative cells more CD34-positive cells were seen in controls than in arthritis patients. .