scholarly journals Retinoic Acid Is a Potent Growth Activator of Mouse Primordial Germ Cells in Vitro

1995 ◽  
Vol 168 (2) ◽  
pp. 683-685 ◽  
Author(s):  
Uichi Koshimizu ◽  
Miho Watanabe ◽  
Norio Nakatsuji
2016 ◽  
Vol 36 (6) ◽  
Author(s):  
Zohreh Makoolati ◽  
Mansoureh Movahedin ◽  
Mehdi Forouzandeh-Moghadam

An in vitro system that supports primordial germ cells (PGCs) survival and proliferation is useful for enhancement of these cells and efficient transplantation in infertility disorders. One approach is cultivation of PGCs under proper conditions that allow self-renewal and proliferation of PGCs. For this purpose, we compared the effects of different concentrations of retinoic acid (RA), and the effect of PGCs co-culture (Co-C) with SIM mouse embryo-derived thioguanine- and ouabain-resistant (STO) cells on the proliferation of embryonic stem cells (ESCs)-derived PGCs. One-day-old embryoid body (EB) was cultured for 4 days in simple culture system in the presence of 5 ng/ml bone morphogenetic protein-4 (BMP4) (SCB group) for PGC induction. For PGC enrichment, ESCs-derived germ cells were cultured for 7 days in the presence of different doses (0–5  μM) of RA, both in the simple and STO Co-C systems. At the end of the culture period, viability and proliferation rates were assessed and expression of mouse vasa homologue (Mvh),  α6 integrin,  β1 integrin, stimulated by retinoic acid 8 (Stra8) and piwi (Drosophila)-like 2 (Piwil2) was evaluated using quantitative PCR. Also, the inductive effects were investigated immunocytochemically with Mvh and cadherin1 (CDH1) on the selected groups. Immunocytochemistry/PCR results showed higher expression of Mvh, the PGC-specific marker, in 3  μM RA concentrations on the top of the STO feeder layer. Meanwhile, assessment of the Stra8 mRNA and CDH1 protein, the specific makers for spermatogonia, showed no significant differences between groups. Based on the results, it seems that in the presence of 3 μM RA on top of the STO feeder layer cells, the majority of the cells transdifferentiated into germ cells were PGCs.


Cell Research ◽  
2021 ◽  
Author(s):  
Xiaoxiao Wang ◽  
Yunlong Xiang ◽  
Yang Yu ◽  
Ran Wang ◽  
Yu Zhang ◽  
...  

AbstractThe pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells (ESCs) and primed epiblast stem cells (EpiSCs), respectively. However, these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development. Here we report the establishment of self-renewing formative pluripotent stem cells (fPSCs) which manifest features of epiblast cells poised for gastrulation. fPSCs can be established from different mouse ESCs, pre-/early-gastrula epiblasts and induced PSCs. Similar to pre-/early-gastrula epiblasts, fPSCs show the transcriptomic features of formative pluripotency, which are distinct from naïve ESCs and primed EpiSCs. fPSCs show the unique epigenetic states of E6.5 epiblast, including the super-bivalency of a large set of developmental genes. Just like epiblast cells immediately before gastrulation, fPSCs can efficiently differentiate into three germ layers and primordial germ cells (PGCs) in vitro. Thus, fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast.


1999 ◽  
Vol 51 (1) ◽  
pp. 208 ◽  
Author(s):  
C-K Lee ◽  
R Weaks ◽  
J.A Piedrahita

2017 ◽  
Vol 45 (7) ◽  
pp. 1608-1619 ◽  
Author(s):  
Kanako Morohaku ◽  
Yuji Hirao ◽  
Yayoi Obata

Author(s):  
Arend W. Overeem ◽  
Yolanda W. Chang ◽  
Jeroen Spruit ◽  
Celine M. Roelse ◽  
Susana M. Chuva De Sousa Lopes

The human germ cell lineage originates from primordial germ cells (PGCs), which are specified at approximately the third week of development. Our understanding of the signaling pathways that control this event has significantly increased in recent years and that has enabled the generation of PGC-like cells (PGCLCs) from pluripotent stem cells in vitro. However, the signaling pathways that drive the transition of PGCs into gonia (prospermatogonia in males or premeiotic oogonia in females) remain unclear, and we are presently unable to mimic this step in vitro in the absence of gonadal tissue. Therefore, we have analyzed single-cell transcriptomics data of human fetal gonads to map the molecular interactions during the sex-specific transition from PGCs to gonia. The CellPhoneDB algorithm was used to identify significant ligand–receptor interactions between germ cells and their sex-specific neighboring gonadal somatic cells, focusing on four major signaling pathways WNT, NOTCH, TGFβ/BMP, and receptor tyrosine kinases (RTK). Subsequently, the expression and intracellular localization of key effectors for these pathways were validated in human fetal gonads by immunostaining. This approach provided a systematic analysis of the signaling environment in developing human gonads and revealed sex-specific signaling pathways during human premeiotic germ cell development. This work serves as a foundation to understand the transition from PGCs to premeiotic oogonia or prospermatogonia and identifies sex-specific signaling pathways that are of interest in the step-by-step reconstitution of human gametogenesis in vitro.


Development ◽  
1996 ◽  
Vol 122 (3) ◽  
pp. 915-923
Author(s):  
W. Halfter ◽  
B. Schurer ◽  
H.M. Hasselhorn ◽  
B. Christ ◽  
E. Gimpel ◽  
...  

A mucin was discovered on the surface of migratory primordial germ cells (PGCs) from chick and rat embryos by means of two monoclonal antibodies. The protein was found to be identical or closely related to ovomucin, a 600 X 10(3) relative molecular mass glycoprotein, and a major constituent of the vitelline membrane of the avian yolk. Based on its resemblance to ovomucin it is referred to as ovomucin-like protein (OLP). The OLP was expressed on PGCs from E3 to E7 female, and from E3 to E12 male chick embryos as the PGCs migrate and colonize the gonadal ridges. After the PGCs have settled in the gonads, they no longer express OLP. In tissue cultures of dissociated cells from E6 gonads, OLP was present only on cells that were positive for PAS staining, the standard histological method to identify PGCs in the chick embryo. Since unfixed PGCs were recognized by the antibodies, at least part of the OLP is localized on the cell surface. The anti-OLP antibodies also stained PGCs in the gonads of the rat embryo, showing that the expression of this antigen on PGCs is phylogenetically conserved. Ovomucin isolated from vitelline membrane prevented adhesion of fibroblasts but not PGCs when used a as a substratum in vitro. The anti-adhesive quality of the mucin resides in the sialic acid residues of the carbohydrate side chains. We propose that OLP has a similar anti-adhesive quality as the ovomucin from vitelline membrane, and that this anti-adhesive property is important to prevent precocious adhesion of migrating PGCs to blood vessel walls and to connective tissue in the mesentery as they migrate toward the gonadal ridges.


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