Differentiation of primordial germ cells from premature ovarian insufficiency-derived induced pluripotent stem cells

Abstract

Background Premature ovarian insufficiency (POI) is a common disease in reproductive women. The pathogenesis of POI is not clear, although it is known that it involves the disorder of oocyte differentiation and development. The introduction of reprogramming human somatic cells into induced pluripotent stem cells (iPSCs) offers a unique opportunity to study many aspects of POI from cell differentiation in vitro that could ultimately lead to novel drug development and testing to help treat the disorder. Methods The fibroblasts from POI patients, including fragile X syndrome, abnormal karyotype (45, X; 45, X/46, XX; 45, XO and 47, XXX), and the gene mutation ( FIGLA and GDF9 ) were reprogrammed to pluripotency status by retroviral transduction using defined factors. The morphology, growth characteristics, gene expression profiles, epigenetic status, and in vitro and in vivo differentiation potential of the POI-1-iPSCs (from fragile X syndrome) were analyzed. Then, POI-1-iPSCs were induced to differentiation into primordial germ cells (PGCs) with DNA methyltransferase inhibitors. Results The iPSCs were successfully generated from POI patients’ fibroblasts. The formed iPS clones have the same characteristics of human ESCs. POI-1-iPSCs were successfully generated with germline competence. The POI-1-iPSCs, with genotypes of fragile X syndrome, can be induced to differentiation into PGCs with high efficiency under our culture system by DNA demethylation. This study proved that disease-specific iPSC lines derived from POI patients could be generated and successfully differentiated into PGCs. Conclusions We established some novel, systemic cell models for the studying of the pathogenesis of POI patients. Second, DNA demethylation may accelerate the induction of human PGCs from iPSCs in vitro and the conclusion needs further exploration. This represents an important step in the novel approach for the study of the pathophysiology and potential egg resource for POI patients. Electronic supplementary material The online version of this article (10.1186/s13287-019-1261-6) contains supplementary material, which is available to authorized users.

Advances in Female Germ Cell Induction from Pluripotent Stem Cells

“…After the researchers demonstrated mouse oocytes from granulosa cell-derived iPSCs possess a higher germline competency than other cell lines, the certain human granulosa cells discarded after IVF were also considered as a more permissive cell source for iPSCs to generate oocytes [ 58 ]. Human iPSCs could provide patient-specific PSCs which could be used to investigate disease-specific pathogenesis in vitro [ 26 , 82 , 83 ]. Recently, a study established human iPSCs in 4i medium from patients with premature ovarian insufficiency.…”

“…Compared to the previous human PGCLCs induction methods, the addition of DNA methyltransferase inhibitor enhanced human PGCLCs induction. Thus, they provided a complementary way for human PGC differentiation from patient-specific iPSCs [ 82 ]. Furthermore, the PSCs pluripotent state was also considered as an important factor during induction.…”

Applying Single-Cell Analysis to Gonadogenesis and DSDs (Disorders/Differences of Sex Development)

“…Organoids can be derived from induced pluripotent stem cells (iPS cells) and can be used to model normal and abnormal development. For the gonad, iPS cells have been used to generate Sertoli-like cells, and also Leydig and germ cells [ 182 , 183 , 184 , 185 ]. Single-cell RNA-seq could be used to chart the differentiation of these cell types, identifying important genes activated during formation of the testis.…”

Pluripotent Cell Models for Gonadal Research

“…In human female iPSCs, Eguizabal et al showed that overexpression of DAZL and BOULE can also lead iPSCs to enter meiosis. Yang et al [105] used fibroblasts from premature ovarian insufficiency (POI) patients to successfully differentiate them into PGCs that had the potential for meiotic progression in vitro. Mimicking the natural role of the oocyte by stimulation with GDF9 and BMP15 proved further induction of folliculogenesis in differentiated PGCs [108].…”