Dataset Information

Establishment and adipocyte differentiation of polycystic ovary syndrome-derived induced pluripotent stem cells.

ABSTRACT:

Objective

To establish a new biological cell model and approach to mimic abnormal lipid metabolism of polycystic ovary syndrome (PCOS) in vitro.

Materials and methods

Epithelial cells from PCOS patients were reprogrammed to pluripotency by retroviral transduction using defined factors. Morphology, growth characteristics, karyotype, gene expression and differentiation in vitro and in vivo were detected by identification protocol of human embryonic stem cells (ESCs). PCOS-induced pluripotent stem cells (iPSCs) were then induced to differentiate into adipocytes. Ability of the adipocytes for glucose consumption was compared with those from non-PCOS-iPSCs.

Results

iPSCs were successfully generated from PCOS patients' adult cells. Formed iPSC clones had the same characteristics of human ESCs. PCOS-iPSCs were induced to differentiation into normal karyotype adipocytes. Compared to non-PCOS-iPSCs, PCOS-iPSCs had more glucose consumption ability during adipocyte differentiation and development in vitro.

Conclusions

This protocol provides a new biological cell model and approach for studying pathogenesis of PCOS and discovering potential drugs to treat it.

SUBMITTER: Yang S

PROVIDER: S-EPMC6496004 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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Publications

Establishment and adipocyte differentiation of polycystic ovary syndrome-derived induced pluripotent stem cells.

Yang Sheng S Ding Shufang S Jiang Xianglong X Sun Bolan B Xu Qianhua Q

Cell proliferation 20160424 3

<h4>Objective</h4>To establish a new biological cell model and approach to mimic abnormal lipid metabolism of polycystic ovary syndrome (PCOS) in vitro.<h4>Materials and methods</h4>Epithelial cells from PCOS patients were reprogrammed to pluripotency by retroviral transduction using defined factors. Morphology, growth characteristics, karyotype, gene expression and differentiation in vitro and in vivo were detected by identification protocol of human embryonic stem cells (ESCs). PCOS-induced pl ...[more]

PMID: 27108524

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Project description:BackgroundPolycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder that affects female fertility. However, with the lack of a corresponding research model, the pathology mechanism of PCOS is poorly understood. Induced pluripotent stem cell (iPSC) technology has been recognized as means to generate patient-specific stem cells for disease modeling.MethodsThe mRNA abundance of iPSCs was analyzed by RNA microarray and real-time polymerase chain reaction (RT-PCR). Karyotyping of iPSCs was performed with cytogenetic analysis. The mitochondrial respiration ability and glycolytic function were measured by the Seahorse Bioscience XF extracellular flux analyzer. The expression of iPSC-associated markers was identified by immunofluorescence and RT-PCR. The teratoma formation of iPSCs was studied using immunochemistry.ResultsA PCOS patient-derived iPSC model was established from somatic cells of PCOS patients. Through comprehensive transcriptional profiling analysis of the RNA microarray, PCOS patient-derived iPSCs showed metabolic abnormalities and mitochondrial dysfunction compared with non-PCOS patient-derived iPSCs in vitro. Specifically, a total of 2904 genes were differentially expressed between the two iPSC populations, of which 1416 genes were upregulated and 1488 genes were downregulated (fold change > 2, p < 0.01). Gene Ontology (GO) term enrichment results showed that upregulated genes were enriched in metabolic processes and mitochondrial activities which participated in the tricarboxylic acid (TCA) cycle, the respiratory electron transport chain (ETC), and glycogenolysis. On the other hand, the downregulated genes were related to cell communication, glucose transport, and uptake. The differentially expressed genes were verified by RT-PCR in PCOS patient-derived iPSCs and granulosa cells from PCOS patients. The PCOS patient-derived iPSCs demonstrated decreased mitochondrial respiration ability and glycolytic function (p < 0.05) but increased mitochondrial copy numbers and biogenesis (p < 0.05). Subsequently, some genes related to glucose metabolism were rescued by treating with metformin in PCOS patient-derived iPSCs. Meanwhile, the ATP production ability of mitochondria and the glycolysis ability of PCOS patient-derived iPSCs also partially returned to normal levels. However, metformin had little effect on mitochondrial maximal respiration ability and maximal glycolytic capacity.ConclusionsWe measured differences in iPSCs from women with and without PCOS in gene transcription and mitochondrial respiratory function. PCOS patient-derived iPSCs showed abnormal expression of metabolic genes and mitochondrial dysfunction in vitro. The study provides a novel cell model in vitro for studying the clinical causes and molecular mechanisms of PCOS.

Dataset Information

Establishment and adipocyte differentiation of polycystic ovary syndrome-derived induced pluripotent stem cells.

Objective

Materials and methods

Results

Conclusions

Publications

Establishment and adipocyte differentiation of polycystic ovary syndrome-derived induced pluripotent stem cells.

Similar Datasets

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