Fibroblasts and transforming growth factor beta induce organization and differentiation of T84 human epithelial cells.
Fibroblasts and transforming growth factor beta induce organization and differentiation of T84 human epithelial cells.
复制标题
成纤维细胞和转化生长因子β诱导T84人上皮细胞的组织和分化。
作者：
T. Halttunen;A. Marttinen;I. Rantala;H. Kainulainen;M. Mäki
背景与目标
肠上皮细胞的隐窝绒毛轴代表了一个连续的发展系统，其中的作用成纤维细胞上皮细胞的相互作用是显而易见的。本研究的目的是建立一种体外方法，从而可以研究成纤维细胞引导的隐窝样肠上皮细胞的分化。
方法
肠上皮细胞（T84和HT-29）在I型胶原凝胶内与成纤维细胞一起培养，没有细胞与细胞接触。T84细胞也在转化生长因子β和肝细胞生长因子的存在下生长。采用光镜、电镜、组织化学和免疫组织化学方法对凝胶进行了研究。
结果
上皮细胞在凝胶内形成无组织的细胞簇，但当给予成纤维细胞支持时，76%的T84细胞集落（不是HT-29）组织成管腔形成，并且包括层粘连蛋白的基底膜沉积良好。柱状单细胞层管腔结构中的细胞（占所有菌落的49%）分化，显示微绒毛、上调的碱性磷酸酶刷状缘活性和小肠典型的粘蛋白谱。这种成纤维细胞诱导的组织化和分化由转化生长因子β诱导。
结论
当与成纤维细胞或转化生长因子β一起三维生长时，隐窝样T84上皮细胞能够分化。该方法可用于间充质-上皮细胞串扰研究。
BACKGROUND & AIMS
The gut epithelium in the crypt-villus axis represents a continuous developmental system in which the role of fibroblast-epithelial interactions is obvious. The aim of this study was to establish an in vitro method whereby fibroblast-guided differentiation of crypt-like gut epithelial cells can be studied.
METHODS
Intestinal epithelial cells (T84 and HT-29) were cultured within type I collagen gel together were fibroblasts without cell-to-cell contact. T84 cells were also grown in the presence of transforming growth factor beta and hepatocyte growth factor. The gels were studied using light and electron microscopy and histochemical and immunohistochemical methods.
RESULTS
The epithelial cells formed unorganized cell clusters within the gels, but when given fibroblast support, 76% of the T84 cell colonies (not HT-29) organized into luminal formations, and basement membranes including laminin were well deposited. The cells in the columnar single cell-layer luminal formations (49% of all colonies) were differentiated, showing microvilli, up-regulated alkaline phosphatase brush border activity, and mucin profiles typical for small intestine. This fibroblast-induced organization and differentiation was induced by transforming growth factor beta.
CONCLUSIONS
Crypt-like T84 epithelial cells are able to differentiate when grown three-dimensionally together with fibroblasts or transforming growth factor beta. This method may be used for mesenchymal-epithelial cell cross-talk studies.