Merne Marina, Syrjänen Stina
Department of Oral Pathology and Radiology, Institute of Dentistry and MediCity Research Laboratory, Faculty of Medicine, University of Turku, Finland.
Arch Dermatol Res. 2003 Sep;295(5):190-8. doi: 10.1007/s00403-003-0419-2. Epub 2003 Aug 1.
Cell-matrix interactions are thought to influence epithelial structure, growth, and differentiation. Three-dimensional cell cultures were used to study the effects of the composition of the dermal equivalent on the morphology of epithelium grown from HaCaT skin keratinocytes. Three commercial preparations, a basement membrane preparation from a tumor (matrix 1), two preparations consisting of collagen type I and III (matrix 3 and 4) and a noncommercial preparation containing collagen type I (matrix 2) were investigated. The effects of fibroblasts of different origin (vaginal mucosa, oral buccal mucosa, and skin) were investigated in connection with matrix 4. The histomorphology and expression of the proteins PCNA, Ki-67, p53, p21, pankeratin, involucrin, cytokeratin 10 (Ck10), Ck17, Ck19 and collagen type IV were evaluated. Three-dimensional cultures of HaCaT cells gave rise to an epithelium with an immature and hyperproliferative character, showing active proliferation with intense PCNA staining. Both matrix 1 and matrix 2 resulted in an epithelium with budding into the matrix and some degree of layering. These epithelia showed only scattered Ck17 and Ck19 expression but a low terminal differentiation potential as indicated by scattered Ck10 and involucrin staining. The epithelia of cocultures with matrices 3 and 4 were positive for Ck17 and Ck19. However, the epithelium on matrix 3 showed strong expression of the terminal differentiation marker Ck10 and involucrin. This methodological study provides evidence of the importance of standardization of the composition of the matrix to avoid confounding effects on epithelial morphology and protein expression in studies using a three-dimensional epithelial culture model.
细胞与基质的相互作用被认为会影响上皮结构、生长和分化。三维细胞培养被用于研究真皮替代物的组成对源自HaCaT皮肤角质形成细胞的上皮形态的影响。研究了三种商业制剂、一种来自肿瘤的基底膜制剂（基质1）、两种由I型和III型胶原蛋白组成的制剂（基质3和4）以及一种含I型胶原蛋白的非商业制剂（基质2）。结合基质4研究了不同来源（阴道黏膜、口腔颊黏膜和皮肤）的成纤维细胞的影响。评估了PCNA、Ki-67、p53、p21、全角蛋白、内披蛋白、细胞角蛋白10（Ck10）、Ck17、Ck19和IV型胶原蛋白的组织形态学及蛋白表达。HaCaT细胞的三维培养产生了具有不成熟和过度增殖特征的上皮，显示出活跃增殖且PCNA染色强烈。基质1和基质2均导致上皮向基质内出芽并出现一定程度的分层。这些上皮仅散在表达Ck17和Ck19，但如散在的Ck10和内披蛋白染色所示，其终末分化潜能较低。与基质3和4共培养的上皮Ck17和Ck19呈阳性。然而，基质3上的上皮显示出终末分化标志物Ck10和内披蛋白的强表达。这项方法学研究提供了证据，证明在使用三维上皮培养模型的研究中，基质组成标准化对于避免对上皮形态和蛋白表达产生混杂效应的重要性。
