Gonadotropins-mediated bovine oocyte in vitro maturation and its genetic and epigenetic change- ART safety in vitro model

Abstract Oocytes of older animals are less likely to be fertilized during the optimal time window post ovulation, resulting in the potential diminished fertilization and embryonic development success. The activity of the epigenetic modifications during this period is a possible target to reverse these damaging effects of aging. The objective of this study was to study the effects of aging during in vitro oocyte maturation in pigs on epigenetic modifications. Oocytes (n = 54) were matured with or without Trichostatin A (TSA; 100 ng/mL), a known meiotic inhibitor, for 24 h, then for an additional 16 h without TSA or hormones for a total of 40 h. At the end of maturation, oocytes were denuded and their zona pellucida’s removed. Oocytes were stained with anti-5-methylcytosine (5mC, 1:500). Fluorescent images of the oocytes were acquired, images were analyzed using ImageJ, and data analysis was performed using ANOVA and Tukey’s test. Oocytes matured with TSA had significantly greater (P < 0.05) levels of DNA methylation by the end of in vitro maturation compared to those not supplemented with TSA These results suggest that TSA can be used to develop an in vitro model to study the effects of epigenetic modifications in oocytes from aged livestock.

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An in vitro model for investigation of vitamin A effects on wound healing

International Journal for Vitamin and Nutrition Research ◽

10.1024/0300-9831/a000692 ◽

2021 ◽

pp. 1-6

Author(s):

Hoda Keshmiri Neghab ◽

Mohammad Hasan Soheilifar ◽

Gholamreza Esmaeeli Djavid

Keyword(s):

Wound Healing ◽

Endothelial Cell ◽

Vitamin A ◽

In Vitro Model ◽

Cellular Proliferation ◽

Endothelial Cell Migration ◽

Normal Fibroblast ◽

Anti Inflammatory ◽

Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inﬂammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inﬂammation responses.

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