Growth of embryonic renal parenchyme at the interphase of a polyester artificial interstitium.
The construction of an artificial kidney module by tissue engineering or the application of cell-based therapies for the treatment of renal failure requires exact information regarding the cellbiological mechanisms of parenchyme development in combination with different kinds of biomaterials. To learn more about these processes tissue cultures are frequently used experimental tools. However, apart from experiments with early kidney anlagen there is a lack of suitable in-vitro models regarding the generation and long-term maintenance of renal tubules. In the present paper we like to demonstrate an advanced culture technique, which allows to generate tubular elements derived from renal stem cells. For the growth of tubules it is essential to fine-tune the interface between the embryonic tissue and the dead fluid space within a perfusion culture container by offering a polyester artificial interstitium. Culture was performed in IMDM supplemented with hormones and growth factors but using serum-free conditions over 14 days. Formation of tissue was then analysed by immunohistochemistry and two-dimensional (2D) electrophoresis. Culture in pure IMDM leads to a complete loss of tissue formation. In contrast, application of aldosterone (A) induces the development of numerous polarised tubules. Surprisingly, addition of epidermal growth factor (EGF), a cocktail of insulin, transferrin and selenium (ITS), retinoic acid (RA), cholecalciferol (VitD3) or bovine pituitary extract (BPT) does not further improve development of tubules, but leads to intensive cell clustering and a decrease of tubule formation. 2D Western blots of developing tissue probed with soybean agglutinin (SBA) reveal a unique pattern of newly detected proteins. It is found that growth factors do not support but abolish protein spots upregulated by aldosterone. It remains to be investigated, which cellbiological effect stimulates the embryonic cells to develop tubules in competition to cell clusters at the interphase of an artificial interstitium.
Minuth
Will W
WW
Department of Molecular and Cellular Anatomy, University of Regensburg, University Street 31, D-93053 Regensburg, Germany. will.minuth@vkl.uni-regensburg.de
Denk
Lucia
L
Heber
Sabine
S
D000450:Aldosterone / Q000737:chemistry; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D015153:Blotting, Western; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D002762:Cholecalciferol / Q000737:chemistry; D046508:Culture Techniques; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D004815:Epidermal Growth Factor / Q000378:metabolism; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D007399:Interphase; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D007684:Kidney Tubules / Q000378:metabolism; D037102:Lectins / Q000737:chemistry; D010477:Perfusion; D010902:Pituitary Gland / Q000378:metabolism; D011091:Polyesters / Q000737:chemistry*; D011506:Proteins / Q000378:metabolism; D011817:Rabbits; D012643:Selenium / Q000378:metabolism; D013499:Surface Properties; D013997:Time Factors; D014168:Transferrin / Q000378:metabolism; D014212:Tretinoin / Q000378:metabolism; D015854:Up-Regulation
D001672:Biocompatible Materials; D007328:Insulin; D037102:Lectins; D011091:Polyesters; D011506:Proteins; D014168:Transferrin; D002762:Cholecalciferol; D000450:Aldosterone; D014212:Tretinoin; D004815:Epidermal Growth Factor; D012643:Selenium