Directed Differentiation of Human Embryonic Stem Cells into Prostate Organoids In Vitro and its Perturbation by Low-Dose Bisphenol A Exposure

Studies using rodent and adult human prostate stem-progenitor cell models suggest that developmental exposure to the endocrine disruptor Bisphenol-A (BPA) can predispose to prostate carcinogenesis with aging. Unknown at present is whether the embryonic human prostate is equally susceptible to BPA du...

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Bibliographic Details
Published in:PloS one 2015-07, Vol.10 (7), p.e0133238
Main Authors: Calderon-Gierszal, Esther L, Prins, Gail S
Format: Article
Language:English
Subjects:
Activin
Adult
Aging
Aging (natural)
Androgens
Animal models
Benzhydryl Compounds - pharmacology
Bisphenol A
Carcinogenesis
Carcinogens
Cell culture
Cell differentiation
Cell Differentiation - drug effects
Cell Proliferation - drug effects
Cells, Cultured
Development and progression
Differentiation
Dose-Response Relationship, Drug
Embryo cells
Embryonic stem cells
Endoderm
Endoderm - cytology
Endoderm - drug effects
Endoderm - metabolism
Estrogens
Exposure
Fibroblast growth factor 10
Fluorescent Antibody Technique
Free Radical Scavengers - pharmacology
Gene expression
Growth
Growth factors
Health aspects
Human Embryonic Stem Cells - cytology
Human Embryonic Stem Cells - drug effects
Human Embryonic Stem Cells - metabolism
Humans
Immunofluorescence
In Vitro Techniques
Male
Mesoderm - cytology
Mesoderm - drug effects
Mesoderm - metabolism
Morphogenesis
Morphogenesis - drug effects
Nests
Noggin protein
Organoids
Organoids - cytology
Organoids - drug effects
Organoids - metabolism
Phenols
Phenols - pharmacology
Progenitor cells
Prostate
Prostate - cytology
Prostate - drug effects
Prostate - metabolism
Prostate cancer
Real-Time Polymerase Chain Reaction
Retinoic acid
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger - genetics
Rodents
Stem cell transplantation
Stem cells
Steroid hormones
Steroids
Testosterone
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Summary:Studies using rodent and adult human prostate stem-progenitor cell models suggest that developmental exposure to the endocrine disruptor Bisphenol-A (BPA) can predispose to prostate carcinogenesis with aging. Unknown at present is whether the embryonic human prostate is equally susceptible to BPA during its natural developmental window. To address this unmet need, we herein report the construction of a pioneer in vitro human prostate developmental model to study the effects of BPA. The directed differentiation of human embryonic stem cells (hESC) into prostatic organoids in a spatial system was accomplished with precise temporal control of growth factors and steroids. Activin-induced definitive endoderm was driven to prostate specification by combined exposure to WNT10B and FGF10. Matrigel culture for 20-30 days in medium containing R-Spondin-1, Noggin, EGF, retinoic acid and testosterone was sufficient for mature prostate organoid development. Immunofluorescence and gene expression analysis confirmed that organoids exhibited cytodifferentiation and functional properties of the human prostate. Exposure to 1 nM or 10 nM BPA throughout differentiation culture disturbed early morphogenesis in a dose-dependent manner with 1 nM BPA increasing and 10 nM BPA reducing the number of branched structures formed. While differentiation of branched structures to mature organoids seemed largely unaffected by BPA exposure, the stem-like cell population increased, appearing as focal stem cell nests that have not properly entered lineage commitment rather than the rare isolated stem cells found in normally differentiated structures. These findings provide the first direct evidence that low-dose BPA exposure targets hESC and perturbs morphogenesis as the embryonic cells differentiate towards human prostate organoids, suggesting that the developing human prostate may be susceptible to disruption by in utero BPA exposures.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0133238