Differentiation of Mouse Primordial Germ Cells into Functional Oocytes In Vitro

Morohaku, Kanako; Hirao, Yuji; Obata, Yayoi

doi:10.1007/s10439-017-1815-7

Cited by 10 publications

(7 citation statements)

References 109 publications

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“…Using a stereomicroscope, we dissected the embryos and collected the gonads attached to the mesonephros, which were removed from the gonad using tungsten needles just before the seeding of the gonad. Our culture method for the ovary was based on the protocol described previously by Morahaku et al 26 , 72 . However, in order to perform in-depth cytological, in situ immunofluorescence, and mitochondrial activity studies, we modified the method of the culture procedure depending on the process to be evaluated—follicular progression, immunofluorescence, or mitochondrial activity.…”

Section: Methodsmentioning

confidence: 99%

The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries

González-Sanz

Barreñada

Rial

et al. 2020

Sci Rep

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Vinclozolin is a pesticide with antiandrogenic activity as an endocrine disruptor compound. Its effects upon the progression of primordial follicles were assessed in cultures of mouse fetal ovaries from the onset of meiotic differentiation of germ cells (13.5 days post coitum) and from both in vivo exposed mice and in vitro exposed ovaries. Exposure of ovaries to vinclozolin—at in vitro dosages ranging from 10 to 200 μM and in 3D ex vivo culture following in vivo exposure to 50 mg/kg bw/day—showed delays in meiocyte differentiation and in follicle growth, even at the lowest in vitro dose exposure. Immunofluorescent analysis showed the presence of the proteins MSY2 and NOBOX in the primary follicles but no difference in the level of protein signals or in the number of follicles in relation to treatment. However, assessing the cytological differentiation of germ cells by detecting the synaptonemal complex protein SYCP3, the exposure to vinclozolin delayed meiotic differentiation from both in vitro- and in vivo-exposed ovaries. These effects were concomitant with changes in the energy metabolism, detected as a relative increase of glycolytic metabolism in live-cell metabolic assays in exposed ovaries.

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Section: Methodsmentioning

confidence: 99%

The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries

González-Sanz

Barreñada

Rial

et al. 2020

Sci Rep

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“…We previously demonstrated that mouse fetal gonads cultured in medium containing fetal bovine serum (FBS) exhibited MOF and/or an absence of follicular compartmentalization despite the development of a large number of growing oocytes. As a result, secondary follicles could not be isolated from these ovaries ( Morohaku, 2019 ; Morohaku et al, 2016 , 2017a ). RNA-seq analysis showed that the estrogen-signaling pathway is excessively activated in in vitro -differentiated ovaries compared with in vivo -derived ovaries before follicle formation.…”

Section: Introductionmentioning

confidence: 99%

Blocking estrogen-induced AMH expression is crucial for normal follicle formation

et al. 2021

Self Cite

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In mammals, primordial follicles assembled in fetuses or during infancy constitute the oocyte resources for life. Exposure to 17beta-estradiol and phytogenic or endocrine-disrupting chemicals during pregnancy and/or the perinatal period leads to the failure of normal follicle formation. However, the mechanisms underlying estrogen-mediated abnormal follicle formation and physiological follicle formation in the presence of endogenous natural estrogen are not well-understood. Here, we reveal that estrogen receptor 1, activated by estrogen, binds to the 5′ region of the anti-Mullerian hormone (Amh) gene and upregulates its transcription before follicle formation in cultured mouse fetal ovaries. Ectopic expression of AMH protein was observed in pregranulosa cells of these explants. Furthermore, AMH addition to the culture medium inhibited normal follicle formation. Conversely, alpha-fetoprotein (AFP) produced in fetal liver reportedly blocks estrogen action, although its role in follicle formation is unclear. We further demonstrated that AFP addition to the medium inhibited ectopic AMH expression via estrogen, leading to successful follicle formation in vitro. Collectively, our in vitro experiments suggest that upon estrogen exposure, the integrity of follicle assembly in vivo is ensured by AFP.

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“…However, there have been no subsequent reports on the production of porcine chimeras. Most iPSC lines fulfilled the criteria of pluripotency, but the cells could not contribute to chimeras or generate cloned piglets [56][57][58][59][60] .…”

Section: Derivation Of Pluripotent Stem Cells From Primordial Germ Cellsmentioning

confidence: 99%

Porcine pluripotent stem cells: progress, challenges and prospects

Han¹,

Miao²,

Hua³

et al. 2019

Front. Agr. Sci. Eng.

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Pluripotent stem cells (PSCs) are characterized by their capacity for high self-renewal and multiple differentiation potential and include embryonic stem cells, embryonic germ cells and induced PSCs. PSCs provide a very suitable model for the studies of human diseases, drugs screening, regenerative medicine and developmental biology research. Pigs are considered as an ideal model for preclinical development of human xenotransplantation, therapeutic approaches and regenerative medicine because of their size and physiological similarity to humans. However, lack of knowledge about the derivation, characterization and pluripotency mechanisms of porcine PSCs hinders progress in these biotechnologies. In this review, we discuss the latest progress on porcine PSCs generation, evaluation criteria for pluripotency, the scientific and technical questions arising from these studies. We also introduce our perspectives on porcine PSC research, in the hope of providing new ideas for generating naive porcine PSCs and animal breeding.

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Differentiation of Mouse Primordial Germ Cells into Functional Oocytes In Vitro

Cited by 10 publications

References 109 publications

The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries

The antiandrogenic vinclozolin induces differentiation delay of germ cells and changes in energy metabolism in 3D cultures of fetal ovaries

Blocking estrogen-induced AMH expression is crucial for normal follicle formation

Porcine pluripotent stem cells: progress, challenges and prospects

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