Heber Sabine, Denk Lucia, Hu Kanghong, Minuth Will W
Department of Molecular and Cellular Anatomy, University of Regensburg, Regensburg, Germany.
Tissue Eng. 2007 Feb;13(2):281-92. doi: 10.1089/ten.2006.0199.
Little information on the structural growth of renal tubules is available. A major problem is the technical limitation of culturing intact differentiated tubules over prolonged periods of time. Consequently, we developed an advanced culture method to follow tubule development. Isolated tissue containing renal progenitor cells was placed in a perfusion culture container at the interphase of an artificial polyester interstitium. Iscove's modified Dulbecco's medium without serum or protein supplementation was used for culture, and the culture period was 13 days. Tissue growth was not supported by addition of extracellular matrix proteins. The development of tubules was registered on cryosections labeled with soybean agglutinin (SBA) and tissue-specific antibodies. Multiple SBA-labeled tubules were found when aldosterone was added to the culture medium. In contrast, culture without aldosterone supplementation displayed completely disintegrated tissue. The development of tubules depended on the applied aldosterone concentration. The use of 1 x 10(-6) M and 1 x 10(-7) M aldosterone produced numerous tubules, while application of 1 x 10(-8) M to 1 x 10(-10) M led to a continuous decrease and finally a loss of tubule formation. The development of labeled tubules in aldosterone-treated specimens took an unexpectedly long period of at least 8 days. The morphogenic effect of aldosterone appeared to be mineralocorticoid hormone-specific since spironolactone and canrenoate abolished the development. Finally, dexamethasone induced widely distributed cell clusters instead of tubules.
关于肾小管结构生长的信息很少。一个主要问题是长时间培养完整的分化肾小管存在技术限制。因此，我们开发了一种先进的培养方法来追踪肾小管的发育。将含有肾祖细胞的分离组织置于人工聚酯间质的界面处的灌注培养容器中。使用不含血清或蛋白质补充剂的伊斯科夫改良杜尔贝科培养基进行培养，培养期为13天。添加细胞外基质蛋白并不能促进组织生长。在用大豆凝集素（SBA）和组织特异性抗体标记的冷冻切片上记录肾小管的发育情况。当向培养基中添加醛固酮时，发现多个SBA标记的肾小管。相比之下，不添加醛固酮的培养物显示组织完全解体。肾小管的发育取决于所应用的醛固酮浓度。使用1×10⁻⁶ M和1×10⁻⁷ M醛固酮可产生大量肾小管，而应用1×10⁻⁸ M至1×10⁻¹⁰ M则导致肾小管形成持续减少并最终丧失。醛固酮处理标本中标记肾小管的发育出乎意料地需要至少8天的长时间。醛固酮的形态发生作用似乎是盐皮质激素特异性的，因为螺内酯和坎利酸钾可消除这种发育。最后，地塞米松诱导广泛分布的细胞簇而不是肾小管。
