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.

ISOLATION AND CULTURE OF FIBROBLASTS FROM ENDOSCOPIC DUODENAL BIOPSIES OF CELIAC PATIENTS / L.r. Roncoroni ; tutor: Maria Teresa Bardella ; coordinatore: Massimo Colombo. Universita' degli Studi di Milano, 2011 Jan 18. 22. ciclo, Anno Accademico 2010.

ISOLATION AND CULTURE OF FIBROBLASTS FROM ENDOSCOPIC DUODENAL BIOPSIES OF CELIAC PATIENTS

L.R. Roncoroni
2011

Abstract

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.
18-gen-2011
Settore BIO/13 - Biologia Applicata
malattia celiaca ; fibroblasti ; colture primarie
BARDELLA, MARIA TERESA
COLOMBO, MASSIMO GIUSEPPE
Doctoral Thesis
ISOLATION AND CULTURE OF FIBROBLASTS FROM ENDOSCOPIC DUODENAL BIOPSIES OF CELIAC PATIENTS / L.r. Roncoroni ; tutor: Maria Teresa Bardella ; coordinatore: Massimo Colombo. Universita' degli Studi di Milano, 2011 Jan 18. 22. ciclo, Anno Accademico 2010.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/150560
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