RESUMO
The epithelium is a highly dynamic system, which plays a crucial role in the homeostasis of the intestinal tract. However, studies on the physiological and pathophysiological functions of intestinal epithelial cells (IECs) have been hampered due to lack of normal epithelial cell models. In the present study, we established a reproducible method for primary culture of mouse IECs, which were isolated from the viable small intestinal crypts of murine fetuses (on embryonic day 19), using type I collagenase and hyaluronidase in a short span of time (≤20 min). With this method, continuously growing mouse IECs, which can be subcultured over a number of passages, were obtained. The obtained cell lines formed a tight cobblestone-like arrangement, displayed long and slender microvilli, expressed characteristic markers (cytokeratin 18 and Notch-1), and generated increasing transepithelial electrical resistance and low paracellular permeability during in vitro culture. The cells also had enzymatic activities of alkaline phosphatase and sucrase-isomaltase, and secreted various cytokines (IL-1β, IL-6, IL-8, and monocyte chemoattractant protein-1), responding to the stimulation of Escherichia coli. These results show that the primary-cultured mouse IECs obtained by the method established here had the morphological and immunological characteristics of IECs. This culture system can be a beneficial in vitro model for studies on mucosal immunology and toxicology.
Assuntos
Animais , Masculino , Feminino , Técnicas de Cultura de Células/métodos , Células Epiteliais/citologia , Hialuronoglucosaminidase , Intestino Delgado/citologia , Metaloproteinase 13 da Matriz , Proliferação de Células , Células Cultivadas , Colagenases , Citocinas/metabolismo , Células Epiteliais/metabolismo , Imunofluorescência , Hematoxilina , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Reprodutibilidade dos Testes , Fatores de TempoRESUMO
Morphea is a rare, sclerotic connective tissue disorder and is thought to be caused by a decreased collagenase activity. Numerous treatment modalities have been tried, such as infiltration with glucocorticosteroid, D-penicillamine, antimalarial agents and cyclosporine. However, all have shown only limited success. We report a case of a 21 year- old female with localized scleroderma, who showed a marked improvement after localized therapy with high dose UVA-1.