Human colon tissue in organ culture: preservation of normal and neoplastic characteristics
Human colon tissue in organ culture: preservation of normal and neoplastic characteristics
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DOI：
10.1007/s11626-009-9247-9
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发表时间：
2010-02-01
影响因子：
2.1
通讯作者：
Varani, James
中科院分区：
文献类型：
--
作者：
Dame, Michael K.;Bhagavathula, Narasimharao;Varani, James
用手术获得的正常和肿瘤人结肠组织建立器官培养的条件。最佳条件为5% CO2和95% O-2的气氛；在气界面处部分组织被粘膜浸没；无血清培养基中添加1.5 mM Ca2+和多种生长补充剂。组织学、组织化学和免疫组织化学特征区分正常和肿瘤组织保存2 d。在正常组织中，这包括存在细长的隐窝，隐窝基部有小而密集的细胞，上部有含有黏液的杯状细胞。增殖细胞的Ki67染色局限于隐窝的下三分之一，而细胞外钙敏感受体的表达见于上三分之一和表面上皮。E-cadherin和β -catenin在整个上皮中表达，并局限于细胞表面。在肿瘤组织中，2 d后出现与时间0时相同的无组织、异常的腺结构。这些结构中的大多数细胞是粘蛋白缺乏的，但偶见杯状细胞和粘蛋白染色。整个异常上皮可见Ki67染色，钙敏感受体表达弱且多变。细胞表面可见e -钙粘蛋白（与正常组织相似），但部分部位可见弥漫性细胞质染色。最后，在培养的肿瘤组织中观察到强烈的细胞质和细胞核β -连环蛋白染色。
Normal and neoplastic human colon tissue obtained at surgery was used to establish conditions for organ culture. Optimal conditions included an atmosphere of 5% CO2 and 95% O-2; tissue partially submerged with mucosa at the gas interface; and serum-free medium with 1.5 mM Ca2+ and a number of growth supplements. Histological, histochemical, and immunohistochemical features that distinguish normal and neoplastic tissue were preserved over a 2-d period. With normal tissue, this included the presence of elongated crypts with small, densely packed cells at the crypt base and mucin-containing goblet cells in the upper portion. Ki67 staining, for proliferating cells, was confined to the lower third of the crypt, while expression of extracellular calcium-sensing receptor was seen in the upper third and surface epithelium. E-cadherin and beta-catenin were expressed throughout the epithelium and confined to the cell surface. In tumor tissue, the same disorganized, abnormal glandular structures seen at time zero were present after 2 d. The majority of cells in these structures were mucin-poor, but occasional goblet cells were seen and mucin staining was present. Ki67 staining was seen throughout the abnormal epithelium and calcium-sensing receptor expression was weak and variable. E-cadherin was seen at the cell surface (similar to normal tissue), but in some places, there was diffuse cytoplasmic staining. Finally, intense cytoplasmic and nuclear beta-catenin staining was observed in cultured neoplastic tissue.