ISOLATION AND CULTURE OF FIBROBLASTS FROM ENDOSCOPIC DUODENAL BIOPSIES OF CELIAC PATIENTS
Introduction and Aim.Esophagogastroduodenoscopy is widely used to investigate the diseases of the upper gastrointestinal tract as celiac disease. Beside its diagnostic and therapeutic potential, it can be useful as source of primary intestinal fibroblasts through the collection of bioptic specimens. Aim of the study is to isolate and culture primary fibroblasts from endoscopic duodenal biopsies of celiac and non-celiac subjects, to analyze their growth patterns and the morphometric characteristics. Patients and Methods. 60 duodenal bioptic specimens from 20 celiac patients and 114 from 38 non-celiac subjects were mechanically chopped and enzymatically digested in order to obtain primary cell cultures. Growth patterns, karyotype (Q-banding analysis), expression of typing proteins (fibroblast surface protein and cytokeratin 20) and morphometric parameters (diameters and their ratio, perimeter, area and perimeter/area ratio at computerised image analysis) were investigated on cultured cells. Results. Primary cells were successfully cultured in 78% of the collected duodenal biopsies. Cultured cells, expressing the fibroblast surface protein, were negative for cytokeratine 20 and maintained a normal kariotype. Cells grew slowly without differences between the celiac and the non celiac group. Morphometric analysis of celiac fibroblasts revealed significantly increased dimensions, with a preserved diameters ratio, and a reduced perimeter/area ratio. Conclusion. For the first time this study demonstrates the feasibility of culturing primary fibroblast cell from endoscopic duodenal biopsies in celiac and non-celiac subjects, opening a new window of opportunity in studies intended to establish the role of fibroblasts as a possible partaker in the pathogenesis of the celiac mucosal damage.
malattia celiaca, fibroblasti, colture primarie, Settore BIO/13 - Biologia Applicata
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