Differentiation of Neonatal Human-Induced Pluripotent Stem Cells to Prostate Epithelial Cells: A Model to Study Prostate Cancer Development
Annual rept. 7 May 2012-6 May 2013
WISCONSIN UNIV MADISON
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In this project we set out to establish conditions for differentiation of human neonatal foreskin skin fibroblast-derived iPSCs into prostate epithelium-like cells and identify differences in gene expression between prostate epithelial cells derived from iPSCs of Caucasian white and African-American black foreskin fibroblasts. We identified and optimized culture conditions that promote prostate epithelial cell-like differentiation of humaniPS clone, IMAR90-4. Our data show that a feeder layer of urogential mesenchymalUGSM cells from neonatal mouse of either gender in combination with neonatal human dermal fibroblasts induced a striking morphological changes that resembles epithelila differentiation with formation of lumen-like structures. We show requirement of components of the extracellular matrix that promote epithelial-type differentaition. Immunofluorescence and biochemivcal analyses showed expression of androgen receptor and markers of epithelial differentiation. Analyses of pluripotency marker expression by RT-PCR showed that while human dermal fibroblasts have higher constitutive expression of Nanog, Oct4 and Sox2 compared to UGSM and IMP90 cells. Preliminary studies also showed that black black fibrobalst population has higher constitutive expression of pluripotency markers than cells from white individuals.
- Medicine and Medical Research