PD35-02 ISOLATION OF NEPHRON PROGENITORS CELLS EXPRESSING SIX2CITED1+ FROM HUMAN FETAL KIDNEYS AND AMNIOTIC FLUID

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2016PD35-02 ISOLATION OF NEPHRON PROGENITORS CELLS EXPRESSING SIX2CITED1+ FROM HUMAN FETAL KIDNEYS AND AMNIOTIC FLUID Stefano Da Sacco, Astgik Petrosyan, Matthew Thornton, Brendan Grubbs, Roger De Filippo, and Laura Perin Stefano Da SaccoStefano Da Sacco More articles by this author , Astgik PetrosyanAstgik Petrosyan More articles by this author , Matthew ThorntonMatthew Thornton More articles by this author , Brendan GrubbsBrendan Grubbs More articles by this author , Roger De FilippoRoger De Filippo More articles by this author , and Laura PerinLaura Perin More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1074AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES In the developing kidney, the formation of new nephrons relies on a small population of self renewing nephrogenic progenitors characterized by the co-expression of Six2 and Cited1. Unfortunately, despite their essential role in the renal formation and maturation, isolation and culture of human nephrogenic progenitor lines has not been successfully achieved and what is known about them is mostly based on rodent models. In this project we report for the first time the isolation of a live cell population co-expressing Six2 and Cited1 from human fetal kidneys and its comparison with a recently subpopulation of Six2+Cited1+ from human amniotic fluid METHODS Six2+ Cited1+ live cells from human amniotic fluid and human fetal kidneys were sorted by using Smartflare RNA probes. RNAseq was performed on positive and negative selections immediately after sorting to characterize their gene expression and evaluate their genetic profile without the confounding effects of cell culture. To prove their potential toward a glomerular fate, differentiation toward podocyte lineage was performed RESULTS Six2+ Cited1+ were successfully isolated by using Smartflare RNA probes from both human fetal kidneys (0.17%) and AF derived hAKPC-P (0.2%). RNASeq of Six2+Cited1+ cell lines confirmed expression, among others, of genes including Six2, Cited1, Osr1, suggesting a nephrogenic signature. Clones and subclones from both populations were derived and expanded for many passages in specific nephrogenic media maintaining Six2 and Cited1 expression. These populations showed the ability to integrate in developing renal structures expressing nephrogenic markers when co-cultured with dissociated/re-aggregated hEK. Moreover, we successfully confirmed differentiation into podocyte-like cells, evaluated by expression of specific markers including WT1 and nephrin, deposition of collagen IV alpha3-4-5 and functional response to angiotensin II and PAN. CONCLUSIONS In conclusion, our preliminary results suggest for the first time, the possibility of deriving and culturing, Six2+Cited1+ cells from human fetal kidneys as well as from an exogenous source of cells like human amniotic fluid, without the use of any genetic manipulation. These systems might represent an accessible and novel source of nephron progenitors that can guide studies of renal cell specification, thus increasing our knowledge of human renal development © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e844 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Stefano Da Sacco More articles by this author Astgik Petrosyan More articles by this author Matthew Thornton More articles by this author Brendan Grubbs More articles by this author Roger De Filippo More articles by this author Laura Perin More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...