Human intestinal goblet cells in monolayer culture: characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line.
Human intestinal goblet cells in monolayer culture: characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line.
复制标题
单层培养中的人肠杯状细胞：源自 HT29 结肠腺癌细胞系的粘液分泌亚克隆的表征。
DOI：
10.1016/0016-5085(88)90678-6
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发表时间：
1988
期刊：
影响因子：
29.4
通讯作者：
Neutra,MR
中科院分区：
文献类型：
--
作者：
Phillips,TE;Huet,C;Bilbo,PR;Podolsky,DK;Louvard,D;Neutra,MR
HT29-18N_2(N_2)细胞是人结肠癌细胞系HT29的亚克隆，是研究人杯状细胞分化和粘蛋白分泌的模型系统。这些细胞在无葡萄糖条件下生长，形成具有典型杯状细胞形态的均质上皮单层柱状细胞。分化发生在未镀膜的玻璃上；层粘连蛋白、纤维连接蛋白或I型或IV型胶原不能促进分化。HT29-18N2细胞生长在无涂层或基质涂层的可渗透滤膜上，形成分化的单层，但其中一些细胞内的粘蛋白颗粒沿上皮腔内极化。针对纯化的人结肠粘蛋白的多克隆抗体，以及针对人结肠粘蛋白的一个蛋白水解酶敏感表位的单抗，将所有分化的杯状细胞的分泌颗粒染色在N_2细胞单层内，但不对缺乏大的分泌颗粒的预分化的杯状细胞进行染色。针对人类粘蛋白特定碳水化合物序列的单抗在分化前也未能对N2细胞进行染色，但识别不同比例的分化后的N2杯状细胞。放射性标记糖蛋白的放射自显影显示了N2细胞粘蛋白颗粒的运输和分泌。胆碱能刺激分化的N_2细胞单层导致细胞内粘蛋白颗粒耗尽。
HT29-18N2 (N2) cells, a subclone of the HT29 human colon carcinoma cell line, are shown in this report to be a model system for the study of human goblet cell differentiation and mucin secretion. Grown in the absence of glucose, these cells formed homogeneous epithelial monolayers of columnar cells with typical goblet cell morphology. Differentiation occurred on uncoated glass; laminin, fibronectin, or collagen type I or IV did not enhance differentiation. HT29-18N2 cells grown on uncoated or matrix-coated permeable filters formed differentiated monolayers, but mucin granules within some of these cells polarized along intraepithelial lumens. Polyclonal antibodies raised against purified human colonic mucin, and also a monoclonal antibody against a protease-sensitive epitope of human colonic mucin, stained secretory granules of all differentiated goblet cells within N2 cell monolayers but did not stain predifferentiated goblet cells lacking large secretory granules. Monoclonal antibodies against specific carbohydrate sequences of human mucins also failed to stain N2 cells before differentiation, but recognized varying fractions of differentiated N2 goblet cells. Autoradiographic visualization of radiolabeled glycoproteins demonstrated transport and secretion of N2 cell mucin granules. Cholinergic stimulation of differentiated N2 cell monolayers resulted in depletion of intracellular mucin granules.