The role of polyester interstitium and aldosterone during structural development of renal tubules in serum-free medium.
Little knowledge is available regarding the development of renal stem/progenitor cells into functional parenchyme. To investigate the environmental mechanisms during this maturation process, we elaborated an advanced culture technique to follow renal tubule development. Embryonic stem/progenitor cells derived from neonatal rabbit kidney were placed in a perfusion culture container at the interphase of an artificial polyester interstitium. Tissue culture was carried out in IMDM without serum or protein supplementation and without coating with extracellular matrix proteins. Development of tubules was registered histochemically on cryosections labeled with soybean agglutinin (SBA) and tissue-specific antibodies. The experiments revealed that the development of renal tubules depends exclusively on the administration of aldosterone. The use of 1x10(-7) M aldosterone for 13 days generated numerous SBA-labeled tubules, while no tubules developed in the absence of the steroid hormone. To obtain further information about the action of the hormone on the cognate receptor, molecular precursors of the aldosterone synthesis pathway were tested. Surprisingly, application of cholesterol, pregnenolone, progesterone, 11-deoxycorticosterone (DOCA) and corticosterone failed to form numerous tubules. Only 11-DOCA and progesterone induced a few tubules, which were barely SBA-labeled. Furthermore, application of aldosterone antagonists such as 1x10(-4) M spironolactone and 1x10(-4) M canrenoate completely inhibited the development of tubules. We conclude that specifically aldosterone promotes the development of tubules via the mineralocorticoid receptor whereas its precursors have no effect.
Minuth
Will W
WW
Department of Molecular and Cellular Anatomy, University of Regensburg, D-93053 Regensburg, Germany. will.minuth@vkl.uni-regensburg.de
Denk
Lucia
L
Hu
Kanghong
K
D000450:Aldosterone / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D000831:Animals, Newborn; D001672:Biocompatible Materials / Q000737:chemistry / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D007684:Kidney Tubules / Q000166:cytology / Q000187:drug effects / Q000196:embryology / Q000254:growth & development; D008954:Models, Biological; D009924:Organ Culture Techniques / Q000379:methods; D037121:Plant Lectins / Q000096:biosynthesis / Q000378:metabolism; D011091:Polyesters / Q000737:chemistry / Q000494:pharmacology*; D011817:Rabbits; D030262:Soybean Proteins / Q000096:biosynthesis / Q000378:metabolism; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods; D010835:Phytohemagglutinins
D000450:Aldosterone; D001672:Biocompatible Materials; D016895:Culture Media, Serum-Free; D037121:Plant Lectins; D011091:Polyesters; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins