Granulosa cell-derived induced pluripotent stem cells exhibit pro-Trophoblastic differentiation potential
Journal
Stem Cell Research and Therapy
Journal Volume
6
Journal Issue
1
Date Issued
2015
Author(s)
Chuang C.-Y.
Huang M.-C.
HSIN-FU CHEN
Yu C.-Y.
Stone L.
Kuo H.-C.
Abstract
Introduction: Human induced pluripotent stem cells (hiPSCs) have been derived from various somatic cell types. Granulosa cells, a group of cells which surround oocytes and are obtained from the (normally discarded) retrieved egg follicles of women undergoing infertility treatment, are a possible cell source for induced pluripotent stem cell (iPSC) generation. Here, we explored the possibility of using human granulosa cells as a donor cell type for iPSC reprogramming, and compared granulosa cell-derived iPSCs (iGRAs) with those derived from other cell sources, to determine the potential ability of iGRA differentiation. Methods: Granulosa cells were collected from egg follicles retrieved from women undergoing infertility treatment. After short-Term culture, the granulosa cells derived from different patients were mixed in culture, and infected with retroviruses encoding reprogramming factors. The resulting iPSC clones were selected and subjected to microsatellite DNA analysis to determine their parental origin. IGRAs were subjected to RT-PCR, immunofluorescence staining, and in vitro and in vivo differentiation assays to further establish their pluripotent characteristics. Results: Microsatellite DNA analysis was used to demonstrate that hiPSCs with different parental origins can be simultaneously reprogrammed by retroviral transfection of a mixed human granulosa cell population obtained from multiple individuals. The iGRAs resemble human embryonic stem cells (hESCs) in many respects, including morphological traits, growth requirements, gene and marker expression profiles, and in vitro and in vivo developmental propensities. We also demonstrate that the iGRAs express low levels of NLRP2, and differentiating iGRAs possess a biased differentiation potential toward the trophoblastic lineage. Although NLRP2 knockdown in hESCs promotes trophoblastic differentiation of differentiating hESCs, it does not result in exit from pluripotency. These results imply that NLRP2 may play a role in regulating the trophoblastic differentiation of human pluripotent stem cells. Conclusions: These findings provide a means of generating iPSCs from multiple granulosa cell populations with different parental origins. The ability to generate iPSCs from granulosa cells not only enables modeling of infertility-Associated disease, but also provides a means of identifying potential clinical interventions through iPSC-based drug screening. ? 2015 Chuang et al.
SDGs
Other Subjects
microsatellite DNA; nucleotide binding oligomerization domain leucine rich repeat and pyrin domain containing protein 2; nucleotide binding protein; unclassified drug; chorionic gonadotropin; estradiol; microsatellite DNA; NLRP2 protein, human; progesterone; signal transducing adaptor protein; small interfering RNA; adult; animal cell; Article; cell clone; cell culture; cell differentiation; cell differentiation assay; cell lineage; cell population; cell structure; controlled study; DNA determination; embryo; embryonic stem cell; female; gene expression profiling; granulosa cell; granulosa cell derived induced pluripotent stem cell; human; human cell; in vitro study; in vivo study; induced pluripotent stem cell; male; mouse; nonhuman; nuclear reprogramming; priority journal; trophoblast; antagonists and inhibitors; cell differentiation; cytology; DNA microarray; fibroblast; genetics; granulosa cell; human embryonic stem cell; induced pluripotent stem cell; metabolism; transplantation; trophoblast; Adaptor Proteins, Signal Transducing; Adult; Cell Differentiation; Cells, Cultured; Cellular Reprogramming; Chorionic Gonadotropin; Estradiol; Female; Fibroblasts; Granulosa Cells; Human Embryonic Stem Cells; Humans; Induced Pluripotent Stem Cells; Male; Microsatellite Repeats; Oligonucleotide Array Sequence Analysis; Progesterone; RNA, Small Interfering; Trophoblasts
Publisher
BioMed Central Ltd.
Type
journal article