Takiguchi Keiko, Yasugi Sadao, Mizuno Takeo
Zoological Institute, Faculty of Science, University of Tokyo, Hongo, Tokyo 113, Japan.
Dev Growth Differ. 1988 Jun;30(3):241-250. doi: 10.1111/j.1440-169X.1988.00241.x.
In vitro organ culture system which permits embryonic chick proventriculus (glandular stomach) to synthesize pepsinogen de novo was developed. Explants of the proventricular rudiment were cultured on Millipore filters in Medium 199 with Earle's salts supplemented with 50% 12-day embryo extract at 38°C in 95% air and 5% CO . In these culture conditions, pepsinogen, a functional marker protein of proventriculus, was first detected after 3 days of cultivation of 6-day chick proventricular rudiment. When recombined and cultured with 6-day proventricular mesenchyme, 6-day oesophageal, proventricular or gizzard (muscular stomach) epithelium expressed pepsinogen while small intestinal epithelium did not. These results were consistent with the previous results obtained by chorioallantoic membrane (CAM) grafting, and showed that the culture conditions are permissive for pepsinogen expression. When recombined and cultured with reaggregated mesenchymal cells isolated from 6-day proventricular mesenchymal fragments, both 6-day proventricular and gizzard epithelia formed glandular structure and expressed pepsinogen. This indicates that the proventricular mesenchymal cells retain the ability to induce morphogenesis and cytodifferentiation of the proventricular epithelium even if the normal organization of proventricular mesenchyme is once destroyed.
开发了一种体外器官培养系统，该系统可使鸡胚前胃（腺胃）从头合成胃蛋白酶原。将前胃原基的外植体置于微孔滤膜上，在含有Earle盐的199培养基中培养，添加50%的12日龄胚胎提取物，于38°C、95%空气和5%二氧化碳的环境中培养。在这些培养条件下，前胃的功能性标记蛋白胃蛋白酶原在6日龄鸡前胃原基培养3天后首次被检测到。当与6日龄前胃间充质重组并培养时，6日龄食管、前胃或肌胃（腺胃）上皮表达胃蛋白酶原，而小肠上皮不表达。这些结果与先前通过绒毛尿囊膜（CAM）移植获得的结果一致，表明该培养条件有利于胃蛋白酶原的表达。当与从6日龄前胃间充质片段分离的重新聚集的间充质细胞重组并培养时，6日龄前胃和肌胃上皮均形成腺结构并表达胃蛋白酶原。这表明，即使前胃间充质的正常组织结构被破坏一次，前胃间充质细胞仍保留诱导前胃上皮形态发生和细胞分化的能力。
