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Int. J. Mol. Sci. 2014, 15(12), 23240-23254; doi:10.3390/ijms151223240

Detection of Abnormal Extracellular Matrix in the Interstitium of Regenerating Renal Tubules

Molecular and Cellular Anatomy, University of Regensburg, University Street 31, D-93053 Regensburg, Germany
These authors contributed equally to this work.
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Received: 16 October 2014 / Revised: 21 November 2014 / Accepted: 8 December 2014 / Published: 15 December 2014
(This article belongs to the Special Issue Artificial Organs)
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Abstract

Stem/progenitor cells are promising candidates for the regeneration of parenchyma in acute and chronic renal failure. However, recent data exhibit that survival of stem/progenitor cells after implantation in diseased renal parenchyma is restricted. To elaborate basic parameters improving survival, cell seeding was simulated under advanced in vitro conditions. After isolation, renal stem/progenitor cells were mounted in a polyester interstitium for perfusion culture. During generation of tubules, chemically defined CO2 Independent Medium or Leibovitz’s L-15 Medium was applied. Specimens were then fixed for transmission electron microscopy to analyze morphological features in generated tubules. Fixation in conventional glutaraldehyde (GA) solution shows development of tubules each exhibiting a polarized epithelium, an intact basal lamina and an inconspicuous interstitium. In contrast, special fixation of specimens in GA solution containing cupromeronic blue, ruthenium red or tannic acid unveils previously not visible extracellular matrix. Control experiments elucidate that a comparable extracellular matrix is not present in the interstitium of the matured kidney. Thus, generation of renal tubules in combination with advanced fixation of specimens for electron microscopy demonstrates that development of abnormal features in the newly developed interstitium has to be considered, when repair of renal parenchyma is performed by implantation of stem/progenitor cells. View Full-Text
Keywords: kidney; stem/progenitor cells; abnormal development; extracellular matrix; glutaraldehyde; cupromeronic blue; ruthenium red; tannic acid; transmission electron microscopy kidney; stem/progenitor cells; abnormal development; extracellular matrix; glutaraldehyde; cupromeronic blue; ruthenium red; tannic acid; transmission electron microscopy
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Minuth, W.W.; Denk, L. Detection of Abnormal Extracellular Matrix in the Interstitium of Regenerating Renal Tubules. Int. J. Mol. Sci. 2014, 15, 23240-23254.

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