&lt;em&gt;In Vitro&lt;/em&gt; Growth of Mouse Preantral Follicles Under Simulated Microgravity

Zhang, Shen; Wu, Yihao; Weng, Yimin; Xu, Zhihui; Chen, Wenmin; Zheng, Dahan; Lin, Weihong; Liu, Jun; Zhou, Ying

doi:10.3791/55641

Cited by 8 publications

(7 citation statements)

References 17 publications

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“…To support the interpretations of the SEM results, the follicles isolated by the enzymatic method were cultured for 24 h. This short-culture has been shown to be an effective procedure in the evaluation of follicular viability as the follicular damage appears only after a few hours, when the follicle returns to its physiological conditions in which case follicular growth does not occur anymore (Vanhoutte et al, 2004). In the present study, peccary PF viability was confirmed using fluorescent probes that confirmed the presence of an intact basal membrane around isolated follicles, similar to that for domestic swine (Ahn et al, 2012), sheep (Lakshminarayana et al, 2014), and mice (Zhang et al, 2017).…”

Section: Discussionsupporting

confidence: 79%

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu)

Campos

Silva

Moreira

et al. 2019

Zygote

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SummaryWe compare the efficiency of mechanical or enzymatic methods, and their combination, for the isolation of ovarian preantral follicles (PFs) from collared peccaries. The ovaries from six females were subjected to the different methods investigated here. For the enzymatic method, ovary fragments were exposed to collagenase type IV in TCM-HEPES medium; the mechanical procedure was based on ovarian cortex dissociation by using a scalpel blade. The residual solution obtained after the mechanical isolation was subjected to the enzymatic procedure. The number of isolated PFs was quantified and classified as primordial, primary, or secondary; their viability was assessed using trypan blue dye assay. To confirm the results, PFs derived from the most efficient method were evaluated for integrity using scanning electron microscopy (SEM) and subjected to a 24 h in vitro culture for subsequent evaluation of viability by using fluorescent probes. A higher number of PFs (P < 0.05) was obtained from the enzymatic method (961.7 ± 132.9) in comparison with the mechanical method (434.3 ± 88.9), but no difference was observed between the two methods and their combination (743.2 ± 92.8). The trypan blue assay showed that the enzymatic method (98.7 ± 0.6%) provided the highest percentage of viable follicles (P < 0.05). Furthermore, SEM confirmed the ultrastructural integrity of the surface architecture of peccary PFs isolated by the enzymatic procedure; epifluorescence microscopy was used to confirm their viability (86.0%). In conclusion, we suggest that the enzymatic method investigated here is useful for the isolation of viable ovarian PFs from collared peccaries.

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Section: Discussionsupporting

confidence: 79%

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu)

Campos

Silva

Moreira

et al. 2019

Zygote

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“…Cultivation in simulated weightlessness of mouse preantral follicles in vitro revealed disruption of microvilli in the zona pellucida , the presence of vacuolized mitochondria and cytoplasmic vacuoles [ 110 ], and a decrease in follicle survival, probably due to aberrant development of granulosa cells [ 111 ]. In another experiment, Kunming mouse oocytes were harvested at the germinal vesicle stage (which is the metaphase of the first meiotic division) by superovulation stimulation.…”

Section: Gametsmentioning

confidence: 99%

Single Cell in a Gravity Field

Ogneva

2022

Life

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The exploration of deep space or other bodies of the solar system, associated with a long stay in microgravity or altered gravity, requires the development of fundamentally new methods of protecting the human body. Most of the negative changes in micro- or hypergravity occur at the cellular level; however, the mechanism of reception of the altered gravity and transduction of this signal, leading to the formation of an adaptive pattern of the cell, is still poorly understood. At the same time, most of the negative changes that occur in early embryos when the force of gravity changes almost disappear by the time the new organism is born. This review is devoted to the responses of early embryos and stem cells, as well as terminally differentiated germ cells, to changes in gravity. An attempt was made to generalize the data presented in the literature and propose a possible unified mechanism for the reception by a single cell of an increase and decrease in gravity based on various deformations of the cortical cytoskeleton.

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“…The reasons for these failures could be related both to the difficulties of conducting an experiment in space flight conditions and to the characteristics of the early embryogenesis of mammals and the maturation of their germ cells. For example, under simulated microgravity conditions by a random positioning machine, a decrease in the survival of mouse follicles was observed with aberrant development of granulosa cells and oocytes, as shown by a reduction in the content of related markers [ 4 ].…”

Section: Introductionmentioning

confidence: 99%

Sperm Motility of Mice under Simulated Microgravity and Hypergravity

Biryukov

Zhdankina

2020

IJMS

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For deep space exploration, reproductive health must be maintained to preserve the species. However, the mechanisms underlying the effect of changes in gravity on male germ cells remain poorly understood. The aim of this study was to determine the effect of simulated micro- and hypergravity on mouse sperm motility and the mechanisms of this change. For 1, 3 and 6 h, mouse sperm samples isolated from the caudal epididymis were subjected to simulated microgravity using a random position machine and 2g hypergravity using a centrifuge. The experimental samples were compared with static and dynamic controls. The sperm motility and the percentage of motile sperm were determined using microscopy and video analysis, cell respiration was determined by polarography, the protein content was assessed by Western blotting and the mRNA levels were determined using qRT-PCR. The results indicated that hypergravity conditions led to more significant changes than simulated microgravity conditions: after 1 h, the speed of sperm movement decreased, and after 3 h, the number of motile cells began to decrease. Under the microgravity model, the speed of movement did not change, but the motile spermatozoa decreased after 6 h of exposure. These changes are likely associated with a change in the structure of the microtubule cytoskeleton, and changes in the energy supply are an adaptive reaction to changes in sperm motility.

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<em>In Vitro</em> Growth of Mouse Preantral Follicles Under Simulated Microgravity

Cited by 8 publications

References 17 publications

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu)

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu)

Single Cell in a Gravity Field

Sperm Motility of Mice under Simulated Microgravity and Hypergravity

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