Abstract: Research into methods of selection and growth of epithelial basal cells is important for its potential use in pediatric surgery. A 3D culture system was used to investigate the behaviour of mesothelial cells present in the wall of human vaginalis peritonei duct (VPD). To this end we used the VPD removed/extracted in the operating theatre from 20 children undergoing surgery for inguinal hernia or hydrocele. Small tissue fragments placed on collagen sponges were cultured for 7, 14 and 21 days in medium supplemented with 10% FBS, and analysed for the expression and distribution of cytokeratins, p63, Ki- 67, vimentin, CD34, and HBME-1. Mesenchymal cells within the stroma were vimentin positive and endothelial cells of small vessels displayed positive staining for CD34. Cytokeratins, p63 and HBME-1 were negative in all stromal cells. In cultured fragments, flat mesothelial cells positive for vimentin, cytokeratins and HBME-1 proliferated, lining the fragment surface and migrating into the sponge. Capillaries showed morphological alterations; however, their immunoreactivity was comparable with the stroma prior to culture. Cells that had migrated into the sponge and displayed characteristics of mesothelial progenitors, predominantly spindle-shaped and stellate, showed heterogeneous expression of markers especially in late phases of cultivation. These cells were constantly positive for vimentin, a small fraction was cytokeratin-positive and a few displayed HBME-1 immunoreactivity. CD34 was found in cells forming small cavities into the matrix, resembling newly formed blood vessels. Further evidence for a mesothelial progenitor comimg from tissue engineering applications suggest that mesothelial cell progenitors are able to switch between different cell phenotypes depending on the local environment. However, only by performing detailed investigations involving selective cell isolation, clonal analysis together with cell labelling and tracking studies, we will begin to determine the true existence of a mesothelial stem cell. Index words: Tissue engineering -Three-dimentional culture - Processus vaginalis - Peritonei - Mesothelial cells
Three dimensional organotypic culture of vaginalis peritonei duct amd expansion of humana mesothelial progenotor cell
SPINELLI, CLAUDIO;
2008-01-01
Abstract
Abstract: Research into methods of selection and growth of epithelial basal cells is important for its potential use in pediatric surgery. A 3D culture system was used to investigate the behaviour of mesothelial cells present in the wall of human vaginalis peritonei duct (VPD). To this end we used the VPD removed/extracted in the operating theatre from 20 children undergoing surgery for inguinal hernia or hydrocele. Small tissue fragments placed on collagen sponges were cultured for 7, 14 and 21 days in medium supplemented with 10% FBS, and analysed for the expression and distribution of cytokeratins, p63, Ki- 67, vimentin, CD34, and HBME-1. Mesenchymal cells within the stroma were vimentin positive and endothelial cells of small vessels displayed positive staining for CD34. Cytokeratins, p63 and HBME-1 were negative in all stromal cells. In cultured fragments, flat mesothelial cells positive for vimentin, cytokeratins and HBME-1 proliferated, lining the fragment surface and migrating into the sponge. Capillaries showed morphological alterations; however, their immunoreactivity was comparable with the stroma prior to culture. Cells that had migrated into the sponge and displayed characteristics of mesothelial progenitors, predominantly spindle-shaped and stellate, showed heterogeneous expression of markers especially in late phases of cultivation. These cells were constantly positive for vimentin, a small fraction was cytokeratin-positive and a few displayed HBME-1 immunoreactivity. CD34 was found in cells forming small cavities into the matrix, resembling newly formed blood vessels. Further evidence for a mesothelial progenitor comimg from tissue engineering applications suggest that mesothelial cell progenitors are able to switch between different cell phenotypes depending on the local environment. However, only by performing detailed investigations involving selective cell isolation, clonal analysis together with cell labelling and tracking studies, we will begin to determine the true existence of a mesothelial stem cell. Index words: Tissue engineering -Three-dimentional culture - Processus vaginalis - Peritonei - Mesothelial cellsI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
