Permissive effect of glutamine on the differentiation of fetal mouse small intestine in organ culture.
Permissive effect of glutamine on the differentiation of fetal mouse small intestine in organ culture.
复制标题
谷氨酰胺对器官培养中胎鼠小肠分化的许可作用。
DOI：
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发表时间：
1985
期刊：
影响因子：
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通讯作者：
R. Calvert
中科院分区：
文献类型：
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作者：
J. Beaulieu;R. Calvert
15日龄小鼠胚胎近三分之一的小肠可在37℃下培养72 h，当使用Trowell-T8培养基时，保持了外植体的完整性，但未形成绒毛，吸收细胞分化差。然而，当使用Leibovitz L-15或RPMI-1640培养基时，可以观察到形成中等大小的绒毛，并且外植体中的吸收细胞分化程度更高。由于T8培养基的化学成分与其他两种培养基有很大的不同，我们决定测试两种主要差异的重要性，即三种氨基酸和五种维生素，以找出哪一种元素是形成肠绒毛所必需的。随后的测试表明，这三种氨基酸对分化的影响是负责的，谷氨酰胺是T8和其他两种培养基之间唯一的关键区别。结果表明，在T8培养基中添加l -谷氨酰胺可使绒毛形成，吸收细胞分化开始，DNA合成增加，最终使上皮细胞数量增加。这些发现表明，只要l -谷氨酰胺在培养基中存在足够的水平，未分化的胎鼠小肠能够在器官培养中表达其表型，即使没有任何外部调节影响。抑制剂的使用表明l -谷氨酰胺可能是葡萄糖胺的能量底物和/或前体所必需的。
The proximal third of the small intestine of 15-day-old mouse embryo can be cultured for 72 h at 37 degrees C. When Trowell-T8 medium is used, the integrity of the explants is maintained, but villi do not form and absorptive cells are poorly differentiated. However, when Leibovitz L-15 or RPMI-1640 medium is used, one can observe the formation of medium-sized villi, and absorptive cells in the explants are more differentiated. Since the chemical composition of T8 medium is quite different from that of the other two media, we decided to test the importance of two major differences, i.e., three amino acids and five vitamins, in order to find out which element(s) is necessary to permit the formation of intestinal villi. Subsequent testing demonstrated that the three amino acids are responsible for the effect on differentiation, and that glutamine is the only critical difference between T8 and the two other media. The results show that the addition of L-glutamine to T8 medium permits the formation of villi, the initiation of absorptive cell differentiation, an increase in DNA synthesis, and finally, an increase in the number of epithelial cells. These findings indicate that undifferentiated fetal mouse small intestine is able to express its phenotype in organ culture, even without any extrinsic regulatory influences, provided that L-glutamine is present at a sufficient level in the culture medium. The use of inhibitors indicated that L-glutamine may be essential as an energetic substrate and/or a precursor for glucosamine.