Gastric parietal cells have a unique secretory membrane system that undergoes a profound transformation when the parietal cell is stimulated to secrete acid. Understanding this process has been hindered by the lack of an immortalised parietal cell line. Here we have explored a strategy for the development of a parietal cell line by the generation of transgenic mice bearing the temperature-sensitive mutant of the SV40 large T antigen (SV40 tsA58) under the control of the regulatory sequences of the gastric H+/K+ ATPase beta-subunit (H/Kbeta-tsA58). Three H/ Kbeta-tsA58 transgenic mouse lines were established, namely 218, 224 and 228, all of which expressed the tsA58 T antigen in the gastric mucosa. Unexpectedly, the gastric mucosae of all lines were hypertrophic indicating that the temperature-sensitive large T antigen was partially active at 37 degrees C. Immunofluorescence together with light and electron microscopic studies revealed that mature parietal and zymogenic cells were absent in H/Kbeta-tsA58 transgenic lines 218 and 224, and small undifferentiated cells were the dominant cell type in the gastric units. On the other hand, a few mature parietal cells were detected in line 228 together with an increased proportion of undifferentiated cells and, normally rare, pre-parietal cells. As line 228 represented a rich source of pre-parietal cells, gastric cells from line 228 were isolated and cultured at 33 degrees C, the permissive temperature for tsA58. Gastric epithelial cells, expressing the T antigen, were maintained in culture for over 6 weeks. Upon a temperature shift to 39 C the cultured gastric cells developed characteristics of differentiated parietal cells, including the presence of a nascent canaliculus and dramatically increased production of the gastric H+/K+ ATPase beta-subunit. Therefore, this system shows the potential to generate an immature parietal cell line that can be induced to differentiate in vitro.

中文翻译：

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表达SV40大T抗原的温度敏感突变体的小鼠胃粘膜摄动。建立永生的壁细胞系的潜力。

胃壁细胞具有独特的分泌膜系统，当刺激壁细胞分泌酸时，胃壁细胞会发生深刻的转化。缺乏永生的壁细胞系阻碍了对这一过程的理解。在这里，我们探索了通过在胃H + / K + ATPase调控序列控制下携带SV40大T抗原（SV40 tsA58）的温度敏感突变体的转基因小鼠的产生来开发壁细胞系的策略。 β亚基（H / Kbeta-tsA58）。建立了三个H / Kbeta-tsA58转基因小鼠品系，即218、224和228，它们均在胃粘膜中表达tsA58 T抗原。不料，所有系的胃粘膜肥大，表明温度敏感的大T抗原在37°C时具有部分活性。免疫荧光以及光镜和电子显微镜研究表明，在H / Kbeta-tsA58转基因系中不存在成熟的壁细胞和酶原细胞218和224，小的未分化细胞是胃单位中的优势细胞类型。另一方面，在品系228中检测到一些成熟的壁细胞，以及未分化细胞和通常罕见的壁前细胞的比例增加。由于线228代表壁前壁细胞的丰富来源，因此分离了线228的胃细胞并在33℃（tsA58的容许温度）下培养。表达T抗原的胃上皮细胞在培养物中维持超过6周。当温度转变为39°C时，培养的胃细胞会形成分化的壁细胞特征，包括新生小管的存在，并显着增加胃H + / K + ATPaseβ亚基的产生。因此，该系统显示了产生不成熟的壁细胞系的潜力，该细胞系可以被诱导在体外分化。