Evaluation of metanephric maturation in a human fetal kidney explant model |
| |
Authors: | Douglas G Matsell Tracey Bennett |
| |
Institution: | (1) Department of Pediatrics, Anatomy and Cell Biology and the Child Health Research Institute, University of Western Ontario, London, Ontario, Canada;(2) Children’s Hospital of Western Ontario, 2230-800 Commissioner’s Road East, N6C 2V5 London, Ontario, Canada |
| |
Abstract: | Summary We have developed a unique human fetal kidney explant model to study the role of the insulinlike growth factor (IGF) system
in metanephric development. Kidneys from 10–18 wk gestation human abortuses were maintained in serum-free conditions and defined
medium, which was shown to support the induction and differentiation of the viable metanephric blastema. Histologically the
tissue remained viable to 192 h of serum-free culture, while metanephric differentiation, reflected by a shrinking nephrogenic
zone and the formation of maturing S-shape and glomerular forms, was accelerated and occurred between 48 and 96 h. In the
nephrogenic zone, a significant decrease in IGF-II gene expression occurred, which reflected the differentiation of the metanephric
blastema cell mass. IGF-II expression persisted, however, in the expanded interstitial mesenchyme. With differentiation over
48 h an increase in IGFBP-2 and WT1 gene expression by Northern blot analysis occurred, and was localized by in situ hybridization to the differentiating glomerular epithelial cell mass. Analysis of the explant-conditioned media by Western
ligand blot demonstrated an increase in the rate of IGF binding protein (IGFBP)-2 peptide production by the differentiating
explant, consistent with an increase in IGFBP-2 gene expression and with metanephric differentiation. This pattern of temporal
and spatial gene expression closely approximates that of normal in vivo fetal renal development and of glomerular epithelial cell differentiation. |
| |
Keywords: | insulinlike growth factor IGF binding protein organ culture human nephrogenesis |
本文献已被 SpringerLink 等数据库收录! |
|