Beaulieu J F, Calvert R
Anat Rec. 1984 Sep;210(1):61-71. doi: 10.1002/ar.1092100110.
The formation of intestinal villi (organogenesis phase) may be studied in organ culture with a completely synthetic medium in 15-day fetal mouse duodenal explants. However, in these explants absorptive cells remained poorly differentiated with all the hormones studied except with epidermal growth factor. In order to elucidate the role of hormones and other factors on the maturation of absorptive cells (maturation phase) in the fetal rodent in organ culture, we have taken the explants after the organogenesis phase. We have studied different culture conditions and have found that 17-day mouse duodenal explants can be cultured during 48 hours with Leibovitz L-15 medium in a 95% O2-5% CO2 atmosphere provided that the explants are relatively large (5 X 2 mm). With this method, dexamethasone (Dx) has been shown to have a direct effect on the maturation of the fetal duodenal mucosa. The addition of Dx (300 ng/ml) to the completely synthetic medium 1) improves the morphology of the explants, 2) induces a significant increase in maltase activity in the tissues, and 3) reduces significantly the labeling index of the duodenal explants after 48 hours of culture. Direct action of Dx on the duodenal mucosa is shown for the first time in organ culture using a completely synthetic medium. This method will permit us to study the effects of other intrinsic and extrinsic factors on the regulation of enzymatic maturation in fetal small intestine.
肠绒毛的形成(器官发生阶段)可以在器官培养中,使用完全合成培养基,以15天胎鼠十二指肠外植体进行研究。然而,在这些外植体中,除了表皮生长因子外,所有研究的激素作用下,吸收细胞的分化仍然很差。为了阐明激素和其他因素对器官培养中胎鼠吸收细胞成熟(成熟阶段)的作用,我们在器官发生阶段之后获取外植体。我们研究了不同的培养条件,发现17天龄小鼠十二指肠外植体,若相对较大(5×2毫米),则可以在含95% O₂ - 5% CO₂的气氛中,用Leibovitz L - 15培养基培养48小时。用这种方法,已证明地塞米松(Dx)对胎鼠十二指肠黏膜的成熟有直接作用。向完全合成培养基中添加Dx(300 ng/ml):1)改善外植体的形态;2)诱导组织中麦芽糖酶活性显著增加;3)在培养48小时后,显著降低十二指肠外植体的标记指数。在使用完全合成培养基的器官培养中,首次显示了Dx对十二指肠黏膜的直接作用。这种方法将使我们能够研究其他内在和外在因素对胎儿小肠酶成熟调节的影响。