Generation of Primordial Germ Cell-like Cells from iPSCs Derived from Turner Syndrome Patients

Aline Fernanda De Souza,Naira Caroline Godoy Pieri,Dimas Tadeu Covas,Tamas Revay,Ramon Cesar Botigelli,Simone Kashima Haddad,Ester Silveira Ramos,Fabiana Fernandes Bressan,Willian Allan King,Flavio Vieira Meirelles

doi:10.3390/cells10113099

Abstract

Turner syndrome (TS) is a genetic disorder in females with X Chromosome monosomy associated with highly variable clinical features, including premature primary gonadal failure leading to ovarian dysfunction and infertility. The mechanism of development of primordial germ cells (PGCs) and their connection with ovarian failure in TS is poorly understood. An in vitro model of PGCs from TS would be beneficial for investigating genetic and epigenetic factors that influence germ cell specification. Here we investigated the potential of reprogramming peripheral mononuclear blood cells from TS women (PBMCs-TS) into iPSCs following in vitro differentiation in hPGCLCs. All hiPSCs-TS lines demonstrated pluripotency state and were capable of differentiation into three embryonic layers (ectoderm, endoderm, and mesoderm). The PGCLCs-TS recapitulated the initial germline development period regarding transcripts and protein marks, including the epigenetic profile. Overall, our results highlighted the feasibility of producing in vitro models to help the understanding of the mechanisms associated with germ cell formation in TS.

Highlights

In humans, Turner syndrome (TS) is caused by complete or partial monosomy of theX chromosome (45, X or mosaic 45, X and 46, XX) [1]
Since the human primordial germ cells (PGCs) from TS patients are challenging to obtain due to ethical procedures and scarce material, other in vitro models, such as the differentiation of induced pluripotent stem cells into primordial germ cell-like cell (PGC-like cells or PGCLCs) can help understand the biological process involved in the pathogenesis of TS
On day 12, all lines of PBMCs-TS and control cell lines enriched with erythroblastic cells were transfected with episomal vectors containing the following pluripotency genes: OCT4, KLF4, SOX2, L-MYC and LIN28 and the other two, p53 and EBNA1, to assist in the reprogramming process (Figure 1A) (Figure S1)

Summary

Introduction

Turner syndrome (TS) is caused by complete or partial monosomy of theX chromosome (45, X or mosaic 45, X and 46, XX) [1]. Turner syndrome (TS) is caused by complete or partial monosomy of the. Previous studies in mice with TS helped the understanding of the patterns of global genomic imbalance involved chromatin regulators and transcriptional regulators in humans [10]. The murine model does not recapitulate the specification of germ cells and primary gonadal failure characteristics of TS patients. Since the human primordial germ cells (PGCs) from TS patients are challenging to obtain due to ethical procedures and scarce material, other in vitro models, such as the differentiation of induced pluripotent stem cells (iPSCs) into primordial germ cell-like cell (PGC-like cells or PGCLCs) can help understand the biological process involved in the pathogenesis of TS

Methods

Results

Discussion

Conclusion