Differentiation of Podocyte and Proximal Tubule-Like Cells from a Mouse Kidney-Derived Stem Cell Line

Abstract

In this study we have shown that the papilla of the mouse kidney contains a population of Pax2+ cells that are detectable from the early postnatal period through to adulthood. Lineage analysis suggests that some of these Pax2+ cells are derived from the metanephric mesenchyme, a population of progenitor cells that gives rise to the nephrons during kidney organogenesis. Here we describe a method for isolating and culturing the Pax2+ population, and demonstrate that some cells within this population are multipotent stem cells, as they are clonogenic and appear to undergo unlimited self-renewal. Further, under appropriate culture conditions, these stem cells can differentiate to generate renal cell types, such as podocyte- and proximal tubule-like cells, and are also able to generate nonrenal cell types, such as adipocytes and osteocytes. The availability of a kidney-derived multipotent stem cell line with the potential to generate podocytes and proximal tubule cells in culture will expedite progress in understanding the biology of these important renal cell types, and will be a useful tool in toxicological studies and drug discovery.

Quantum Dots Do Not Affect the Behaviour of Mouse Embryonic Stem Cells and Kidney Stem Cells and Are Suitable for Short-Term Tracking

“…qPCR analysis of EBs at 7 days showed that expression levels of the endoderm-specific gene, Gata6 [23], the mesoderm-specific gene, brachyury [24] and the ectoderm-specific gene, Pax6 [25], were not significantly different in EBs generated from QD-labelled ESCs compared to controls (Figure 2E). To investigate the effect of QDs on KSC differentiation, qPCR analysis showed that mRNA levels of the Wt1 KSC marker [16] were not significantly different between QD-labelled KSCs and controls (Figure 3A). Furthermore, the majority of QD-labelled cells displayed Wt1 immunoreactivity (Figure 3B).…”

“…The cells were cultured in advanced high glucose DMEM (Invitrogen, UK) supplemented with 2% FCS (PAA laboratories, UK), 2 mM L-glutamine (Sigma, UK) and 0.01% (v/v) 50 mM 2-mercaptoethanol (Invitrogen) on plastic tissue culture dishes (Nunc, Denmark) coated with 0.1% (w/v) gelatine (Sigma). Mouse KSCs were generated by Cristina Fuente Mora from mouse neonatal kidneys in our lab [16]. To generate EGFP + cells (KSC-GFP), KSC cells were transduced with an EGFP-expressing lentivirus under the control of the spleen focus-forming virus (SFFV) promoter, pseudo-coated with a vesicular-stomatitis-virus glycoprotein (VSV-G) envelope.…”

“…To this end, we examined the effect of QDs on the viability, proliferation rate and differentiation potential of two types of stem cells: mouse embryonic stem cells and mouse kidney-derived stem cells (KSCs), a tissue-specific stem cell line isolated from postnatal mouse kidney [16]. We also examined the extent to which QDs are depleted from these stem cells as they proliferate in culture, and determined if QDs released from living or dead cells can be transferred to unlabelled neighbouring cells.…”

MS-1magA

“…Multipotent murine mesenchymal stem/stromal cells (CRL-12424; ATCC, Teddington, United Kingdom), mouse kidney-derived stem cells H6, 22 and HEK 293 T(N) cells (LV900A-1; System Biosciences, Mountain View, California) were cultured in Dulbecco Modified Eagle Medium (DMEM) containing 10% fetal calf serum (FCS) and 1% l -glutamine at 37°C under a humidified atmosphere with 5% CO 2 . All culture media and supplements were purchased from Sigma-Aldrich, Gillingham, United Kingdom, unless stated otherwise.…”

CD117+amniotic fluid stem cells

“…Therefore, the possibility of developing stem cell-based therapies for both glomerular and tubular repair is receiving intensive investigation. 75 Different stem cell types have shown some potential in the generation of functional nephrons [76][77][78][79][80][81] but the most appropriate cell type for transplantation is still to be established. 82 The first evidence that AFS cells could be of relevance for renal stem cell therapy has been published in 2007.…”