Perin First hour 1

The most important demonstraon of the pluripotenality

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Unformatted text preview: atory Germinal layer forma&on Origin of the kidney Intermediate mesoderm - located dorsolaterally, between the somites and the lateral plates - gives rise to the urogenital system, the kidneys and the reproduc&ve organs Metanephros Interac&ons between MM and ureteric bud MM induces elongation and branching of bud MM Bud tips induce MM aggregation Nephric duct Ureteric bud elongates into MM Presence of bud stops MM PCD - Metanephric Mesenchyme and Ureteric Bud exchange signals (GDNF FGF) - MM condensa&on Ureteric Bud branching and consequent epithelialisa&on of MM cells - Following the invagina&on of the epithelialised cells we have the forma&on of C- and S-shapes and the construc&on of vascular elements from blood progenitors - Tubular matura&on Podocyte Endothelial cell Tissue engineering Stem Cell Ingredients for Renal Tissue Engineering in our laboratory: 1. Func&onal cells: stem cells (AFSC). 2. Obtain the suitable scaffold (embryonic kidney or natural matrix). 3. Evaluate if stem cells can survive when seeded into the scaffold. 4. Determine if "created structure" can be implantable in an animal model. 5. Finally our future direc&on is to obtain func&onal renal &ssue Embryonic kidney and AFSC Objec=ve To determine the capacity of the AFSC, once injected into embryonic kidney in vitro, to survive, replicate, integrate themselves into normal kidney. In vitro experiment Cells injec=on 4x Lac-Z 40x CM-Dil 40x GFP 40x In vitro Results Perin L. et al., 2007 Cell Prolifera&on Control GDNF Zo-1 9 days 10x c-shape body Cla ac=n S-shape body E 13, 3 days co-culture, Lac-Z, H&E 20x Live imaging 1 day aKer injec&on 10 days aKer injec&on Live Microscopy, GFP-cells, 4 days Under fluorescent microscopy you can see the green labelled stem cells within the kidney, but more importantly live microscopy demonstrated that these cells con&nued to divide and proliferate under culture condi&ons incorpora&ng themselves within the organ. AKer 10 days, the cells ha...
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