Stage-specific tissue and cell interactions play key roles in mouse germ cell specification

Development ◽  
2001 ◽  
Vol 128 (4) ◽  
pp. 481-490
Author(s):  
T. Yoshimizu ◽  
M. Obinata ◽  
Y. Matsui
Keyword(s):  
Primordial Germ Cells ◽  
Cell Interactions ◽  
Cell Specification ◽  
In Vitro System ◽  
Tissue Interactions ◽  
Germ Cell Specification ◽  
Specific Tissue ◽  
Specific Number ◽  
Embryonic Ectoderm

Primordial germ cells (PGCs) in mice have been recognized histologically as alkaline phosphatase (AP) activity-positive cells at 7.2 days post coitum (dpc) in the extra-embryonic mesoderm. However, mechanisms regulating PGC formation are unknown, and an appropriate in vitro system to study the mechanisms has not been established. Therefore, we have developed a primary culture of explanted embryos at pre- and early-streak stages, and have studied roles of cell and/or tissue interactions in PGC formation. The emergence of PGCs from 5.5 dpc epiblasts was observed only when they were co-cultured with extra-embryonic ectoderm, which may induce the conditions required for PGC formation within epiblasts. From 6.0 dpc onwards, PGCs emerged from whole epiblasts as did a fragment of proximal epiblast that corresponds to the area containing presumptive PGC precursors without neighboring extra-embryonic ectoderm and visceral endoderm. Dissociated epiblasts at these stages, however, did not give rise to PGCs, indicating that interactions among a cluster of a specific number of proximal epiblast cells is needed for PGC differentiation. In contrast, we observed that dissociated epiblast cells from a 6.5-b (6.5+15-16 hours) to 6.75 dpc embryo that had undergone gastrulation gave rise to PGCs. Our results demonstrate that stage-dependent tissue and cell interactions play key roles in PGC determination.

scholarly journals Resf1 supports embryonic stem cell self-renewal and effective germline entry

2021 ◽  
Author(s):  
Matus Vojtek ◽  
Ian Chambers
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Germ Line ◽  
Primordial Germ Cell ◽  
Embryonic Stem ◽  
Cell Specification ◽  
Self Renewal ◽  
Downstream Target ◽  
Germ Cell Specification

Retroelement silencing factor 1 (Resf1) interacts with the key regulators of mouse embryonic stem cells (ESCs) Oct4 and Nanog, and its absence results in sterility of mice. However, the function of Resf1 in ESCs and germ line specification is poorly understood. In this study, we used Resf1 knockout cell lines to determine the requirements of RESF1 for ESCs self-renewal and for in vitro specification of ESCs into primordial germ cell-like cells (PGCLCs). We found that deletion of Resf1 in ESCs cultured in serum and LIF reduces self-renewal potential whereas episomal expression of RESF1 has a modest positive effect on ESC self-renewal. In addition, RESF1 is not required for the capacity of NANOG and its downstream target ESRRB to drive self-renewal in the absence of LIF. However, Resf1 deletion reduces efficiency of PGCLC differentiation in vitro. These results identify Resf1 as a novel player in the regulation of pluripotent stem cells and germ cell specification.

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

Cells ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 3099
Author(s):  
Aline Fernanda de Souza ◽  
Fabiana Fernandes Bressan ◽  
Naira Caroline Godoy Pieri ◽  
Ramon Cesar Botigelli ◽  
Tamas Revay ◽  
...  
Keyword(s):  
Germ Cell ◽  
Turner Syndrome ◽  
Genetic Disorder ◽  
Primordial Germ Cells ◽  
Primordial Germ Cell ◽  
Cell Formation ◽  
In Vitro Models ◽  
Germline Development ◽  
Germ Cell Specification

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.

Adhesiveness of mouse primordial germ cells to follicular and Sertoli cell monolayers

Development ◽  
1985 ◽  
Vol 87 (1) ◽  
pp. 87-97
Author(s):  
Massimo De Felici ◽  
Gregorio Siracusa
Keyword(s):  
Somatic Cell ◽  
Germ Cells ◽  
Primordial Germ Cells ◽  
Gonadal Development ◽  
Cellular Interactions ◽  
Follicular Cells ◽  
Vitro Assay ◽  
In Vitro System ◽  
Cell Monolayers

The adhesiveness of female and male mouse primordial germ cells (PGCs) to somatic cell monolayers of various origin has been studied in the definite conditions of an in vitro system. PGCs were isolated from the gonads of embryos of various post coital ages according to the method of De Felici & McLaren (1982), and seeded on the cell monolayers. PGCs from 12·5 to 15·5 days post coitum (dpc) embryos specifically adhered to Sertoli and follicular cells obtained from adult gonads. The percentage of female PGCs which adhered to follicular cell monolayers was significantly higher than that of male PGCs. No significant adhesion was seen between PGCs and somatic cell monolayers obtained from various embryonic and adult tissues. The results obtained indicate that the simple in vitro assay described in the present paper might help to characterize the cellular interactions between somatic and germ cells during gonadal development.

Strain-dependent viral dynamics and virus-cell interactions in a novel in vitro system supporting the life cycle of blood-borne hepatitis C virus

Hepatology ◽  
2009 ◽  
Vol 50 (3) ◽  
pp. 689-696 ◽  
Author(s):  
Hussein Hassan Aly ◽  
Yue Qi ◽  
Kimie Atsuzawa ◽  
Nobuteru Usuda ◽  
Yasutsugu Takada ◽  
...  
Keyword(s):  
Life Cycle ◽  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Cell Interactions ◽  
Viral Dynamics ◽  
In Vitro System ◽  
Strain Dependent

scholarly journals Complete in vitro generation of fertile oocytes from mouse primordial germ cells

2016 ◽  
Vol 113 (32) ◽  
pp. 9021-9026 ◽  
Author(s):  
Kanako Morohaku ◽  
Ren Tanimoto ◽  
Keisuke Sasaki ◽  
Ryouka Kawahara-Miki ◽  
Tomohiro Kono ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Regenerative Medicine ◽  
Genomic Imprinting ◽  
Germ Cells ◽  
Culture System ◽  
Primordial Germ Cells ◽  
Oocyte Growth ◽  
In Vitro System ◽  
In Vitro Reconstitution

Reconstituting gametogenesis in vitro is a key goal for reproductive biology and regenerative medicine. Successful in vitro reconstitution of primordial germ cells and spermatogenesis has recently had a significant effect in the field. However, recapitulation of oogenesis in vitro remains unachieved. Here we demonstrate the first reconstitution, to our knowledge, of the entire process of mammalian oogenesis in vitro from primordial germ cells, using an estrogen-receptor antagonist that promotes normal follicle formation, which in turn is crucial for supporting oocyte growth. The fundamental events in oogenesis (i.e., meiosis, oocyte growth, and genomic imprinting) were reproduced in the culture system. The most rigorous evidence of the recapitulation of oogenesis was the birth of fertile offspring, with a maximum of seven pups obtained from a cultured gonad. Moreover, cryopreserved gonads yielded functional oocytes and offspring in this culture system. Thus, our in vitro system will enable both innovative approaches for a deeper understanding of oogenesis and a new avenue to create and preserve female germ cells.

scholarly journals Differential repression of Otx2 underlies the capacity of NANOG and ESRRB to induce germline entry

2021 ◽  
Author(s):  
Matus Vojtek ◽  
Jingchao Zhang ◽  
Juanjuan Sun ◽  
Man Zhang ◽  
Ian Chambers
Keyword(s):  
Transcription Factor ◽  
Germ Cells ◽  
Target Gene ◽  
Primordial Germ Cells ◽  
The Core ◽  
Underlying Mechanisms ◽  
Post Implantation ◽  
Embryonic Ectoderm

Primordial germ cells (PGCs) are induced in the embryo by signals, including BMP emanating from extra-embryonic ectoderm, that act on cells in the post-implantation epiblast. PGC development can be recapitulated in vitro through the exposure of epiblast-like cells (EpiLCs) to appropriate cytokines, resulting in differentiation into PGC-like cells (PGCLCs). Interestingly, the requirement for cytokines to induce PGCLCs can be bypassed by enforced expression of the transcription factor (TF) NANOG. However, the underlying mechanisms are not fully elucidated. Here, we show that Otx2 downregulation is essential to enable NANOG to induce PGCLC formation. Moreover, while previous work has shown that the direct NANOG target gene Esrrb can substitute for several functions of NANOG, enforced expression of ESRRB cannot promote PGCLC specification from EpiLCs. This appears to be due to differential downregulation of Otx2 by NANOG and ESRRB, since induction of ESRRB in Otx2+/- EpiLCs activates expression of the core PGC TFs, Blimp1, Prdm14 and Ap2γ and emergence of PGCLCs. This study illuminates the interplay of TFs occurring at the earliest stages of PGC specification from a state of competence.

scholarly journals A Novel Three-Dimensional In Vitro System to Study Trophoblast?Endothelium Cell Interactions

2007 ◽  
Vol 58 (2) ◽  
pp. 98-110 ◽  
Author(s):  
Paulomi B. Aldo ◽  
Graciela Krikun ◽  
Irene Visintin ◽  
Charles Lockwood ◽  
Roberto Romero ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Interactions ◽  
In Vitro System

scholarly journals Comparative Principles of DNA Methylation Reprogramming during Human and Mouse In Vitro Primordial Germ Cell Specification

2016 ◽  
Vol 39 (1) ◽  
pp. 104-115 ◽  
Author(s):  
Ferdinand von Meyenn ◽  
Rebecca V. Berrens ◽  
Simon Andrews ◽  
Fátima Santos ◽  
Amanda J. Collier ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Germ Cell ◽  
Primordial Germ Cell ◽  
Cell Specification ◽  
Germ Cell Specification ◽  
Human And Mouse
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