Construction and characterization of a multilayered gingival keratinocyte culture model: the TURK-U model

摘要: In construction of epithelial cells as multilayers, the are grown submerged to confluence on fibroblast-embedded collagen gels and, then, lifted air promote their stratification. We recently demonstrated that gingival form uniform monolayers semi-permeable nitrocellulose membranes, supported with a semi-solid growth medium, which allows grow at an air–liquid–solid interface from beginning culturing protocol. this study, aim was further develop our previous model multilayered culture model. Two different cell lines (HaCaT skin and HMK gingiva) were used in all experiments. Both first for 3 days. After that, keratinocytes trypsinized, counted seeded sterile membrane placed top forming grow. At days 1, 4, 7, fixed, embedded paraffin, sectioned routine Hematoxylin-Eosin staining immunohistochemistry cytokeratin (Ck). day grew monolayers, while HaCaT stratified epithelium two three layers. had four five layers its proliferation continued up 7. formed dense weakly proliferating stratification 4 but disappeared days, both models strongly positive Ck5, Ck7, Ck 19, Ck10. Gingival stratify successfully medium. This technique researchers construct multilayers interface, without using gels, resulting more reproducible method.