A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues
A method for establishing human primary gastric epithelial cell culture from fresh surgical gastric tissues
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一种从手术新鲜胃组织中建立人原代胃上皮细胞培养物的方法
DOI：
10.3892/mmr.2015.3692
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发表时间：
2015-08-01
影响因子：
3.4
通讯作者：
Yan, Qiu
中科院分区：
文献类型：
--
作者：
Aziz, Faisal;Yang, Xuesong;Yan, Qiu
目前，胃癌的研究和分析主要采用活检标本、胃癌细胞系和胃手术组织，包括胃癌发生的分子机制和蛋白质组学研究。然而，成纤维细胞和其他组织成分可能会干扰这些技术。因此，本研究旨在开发一种从胃手术组织中分离活的人胃上皮细胞的方法。开发了一种使用新鲜手术切除的组织培养人胃上皮细胞的方法，并使用免疫细胞化学、过碘酸-希夫（PAS）染色和细胞活力测定进行了评价。在组织外植体培养的第七天，在胃组织周围观察到低细胞生长。细胞生长随后增加，并且在外植体后12天检测到大量的纯上皮细胞。胃癌细胞生长迅速，倍增时间为13-52 h，而正常细胞的倍增时间为20-53 h。原代胃细胞的免疫细胞化学分析显示细胞角蛋白18和19染色阳性，这表明培养物由纯上皮细胞组成，不含成纤维细胞。PAS染色显示培养的胃细胞产生中性粘蛋白。颗粒蛋白和碳水化合物抗原724染色证实培养的胃癌和正常细胞的纯度。这种细胞培养方法表明原代培养的胃细胞由分泌粘蛋白的胃上皮细胞组成，这可能有助于研究胃幽门螺杆菌感染和胃癌。
At present, biopsy specimens, cancer cell lines and tissues obtained by gastric surgery are used in the study and analysis of gastric cancer, including the molecular mechanisms and proteomics. However, fibroblasts and other tissue components may interfere with these techniques. Therefore, the present study aimed to develop a procedure for the isolation of viable human gastric epithelial cells from gastric surgical tissues. A method was developed to culture human gastric epithelial cells using fresh, surgically excised tissues and was evaluated using immunocytochemistry, periodic acid-Schiff (PAS) staining and cell viability assays. Low cell growth was observed surrounding the gastric tissue on the seventh day of tissue explant culture. Cell growth subsequently increased, and at 12 days post-explant a high number of pure epithelial cells were detected. The gastric cancer cells exhibited rapid growth with a doubling time of 13-52 h, as compared to normal cells, which had a doubling time of 20-53 h. Immunocytochemical analyses of primary gastric cells revealed positive staining for cytokeratin 18 and 19, which indicated that the culture was comprised of pure epithelial cells and contained no fibroblasts. Furthermore, PAS staining demonstrated that the cultured gastric cells produced neutral mucin. Granulin and carbohydrate antigen 724 staining confirmed the purity of gastric cancer and normal cells in culture. This method of cell culture indicated that the gastric cells in primary culture consisted of mucin-secreting gastric epithelial cells, which may be useful for the study of gastric infection with Helicobacter pylori and gastric cancer.