Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro

Paulino, Laís R F M; Barroso, P.A.A.; Silva, A.W.B.; Souza, Ana Liza Paz; Bezerra, F.T.G.; Silva, B.R.; Donato, Mariana M.A.; Peixoto, Christina A.; Silva, J.R.V.

doi:10.1016/j.theriogenology.2019.10.031

Cited by 9 publications

(4 citation statements)

References 40 publications

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“…It has been demonstrated stem cells secrete a variety of cytokines and growth factors, such as EGF, IL, LIF, FGF2, TGF-β, and IGF-1, that regulate the physiological conditions of recipient cells [33][34][35][36]. The beneficial effects of these factors on IVM have been demonstrated in several species, such as human [37,38], cow [39], horse [40], pig [41], and mouse [38]. In the present study, it was demonstrated that EPC can secrete various paracrine factors, including bFGF, VEGF, IGF-1, and IL-10, into the extracellular environment, which is consistent with previous findings [42][43][44].…”

Section: Discussionmentioning

confidence: 99%

Effects of Human Endothelial Progenitor Cell and Its Conditioned Medium on Oocyte Development and Subsequent Embryo Development

Lee

2020

IJMS

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Human endothelial progenitor cells (EPCs) secrete numerous growth factors, and they have been applied to regenerative medicine for their roles in angiogenesis as well as neovascularization. Angiogenesis is one of the essential factors for the maturation of ovarian follicles; however, the physiological function of EPCs or their derivatives on in vitro culture systems has not been fully understood. The aim of this study was to evaluate the effectiveness of EPCs and their conditioned medium (EPC-CM) on oocyte development and subsequent embryo development. In the results, the oocyte development and subsequent embryo development were significantly improved in EPCs and the EPC-CM group. In addition, markedly increased levels of growth factors/cytokines, such as basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), insulin growth factor-1 (IGF-1), interleukin-10 (IL-10), and epidermal growth factor (EGF), were observed in medium from the EPC-CM group. Additionally, EPC-CM after in vitro maturation (IVM) had significantly decreased reactive oxygen species (ROS) levels compared to those of other groups. Transcriptional levels of growth factor receptor-related genes (FGFR2, IGF1R) and anti-apoptotic-related gene (BCL2) were significantly upregulated in cumulus cells/oocytes from the EPC-CM group compared with those from the control. Furthermore, the expression levels of cumulus expansion-related genes (PTGS2, TNFAIP6, HAS2) and oocyte-maturation-related factors (GDF9, BMP15) were significantly enhanced in the EPC-CM group. Consequently, the present study provides the first evidence that EPC-CM contains several essential growth factors for oocyte development by regulating genes involved in oocyte maturation.

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Section: Discussionmentioning

confidence: 99%

Effects of Human Endothelial Progenitor Cell and Its Conditioned Medium on Oocyte Development and Subsequent Embryo Development

Lee

2020

IJMS

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“…Subsequently secondary follicles (~0.2 mm) were visualized under a stereomicroscopic microscope (SMZ 645 Nikon, Tokyo, Japan; ×40 magnification) and dissected manually from fragments using 25G needles. Follicles with visible oocytes surrounded by granulosa cells, without an antral cavity and with an intact basement membrane were selected for culture (Paulino et al, 2018(Paulino et al, , 2020. Follicles with an extruded oocyte and opaque granulosa cells were considered to be atretic (Paulino et al, 2018(Paulino et al, , 2020.…”

Section: Isolation and In Vitro Culture Of Preantral Folliclesmentioning

confidence: 99%

“…Follicles with visible oocytes surrounded by granulosa cells, without an antral cavity and with an intact basement membrane were selected for culture (Paulino et al, 2018(Paulino et al, , 2020. Follicles with an extruded oocyte and opaque granulosa cells were considered to be atretic (Paulino et al, 2018(Paulino et al, , 2020. After selection, the follicles were cultured individually in drops of medium under mineral oil in Petri dishes (60 × 15 mm, Corning, USA).…”

Section: Isolation and In Vitro Culture Of Preantral Folliclesmentioning

confidence: 99%

“…Follicles with opaque granulosa cells and/or darkened oocytes, as well as damaged basal membranes were considered atretic. The formation of the antrum was determined by visualizing a translucent cavity between the layers of granulosa cells (Paulino et al, 2020). The follicle diameter (μm) was calculated only in morphologically normal follicles using two perpendicular measurements from the photographic records of follicles and using an inverted microscope with NIS Elements 2.4 software (Nikon, Nikon Instruments Inc., Japan).…”

Section: Evaluation Of Morphology and Development Of Cultured Folliclesmentioning

confidence: 99%

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Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro

Barroso

Paulino

Silva

et al. 2020

Zygote

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Summary This study aimed to evaluate the effects of dexamethasone on development, viability, antrum formation and ultrastructural integrity of bovine secondary follicles cultured in vitro for 18 days. Bovine ovaries were obtained from slaughterhouses and secondary follicles of ~150–200 µm diameter were isolated and cultured in the laboratory in TCM-199+ alone or supplemented with different concentrations of dexamethasone (1, 10, 100 and 1000 ng/ml). Follicle viability was evaluated after the culture period, using calcein-AM (viable) and ethidium homodimer (nonviable). Follicle diameters and antrum formation were evaluated at days 0, 6, 12 and 18. Before or after in vitro culture, follicles were fixed for histological and ultrastructural analysis. Follicle diameters were evaluated using analysis of variance and Kruskal–Wallis test, while chi-squared test was used to evaluate the percentage of viable follicles and antrum formation (P < 0.05). Follicles cultured for 6 days with all treatments increased their diameters significantly, but there was no significant difference between treatments at the end of the culture period. In vitro cultured follicles showed antral cavity formation at the end of the culture period, but no influence of dexamethasone was seen. Ultrastructural analysis showed that follicles cultured with dexamethasone (1, 10, 100 and 1000 ng/ml) had well preserved granulosa cells. However, oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone showed signs of degeneration. It can be concluded that follicles cultured in vitro in the presence of dexamethasone demonstrated continuous in vitro growth, but oocytes from follicles cultured with 10, 100 or 1000 ng/ml dexamethasone had poor ultrastructure.

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Why Is It So Difficult To Have Competent Oocytes from In vitro Cultured Preantral Follicles?

et al. 2022

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Effects of epidermal growth factor and progesterone on development, ultrastructure and gene expression of bovine secondary follicles cultured in vitro

Cited by 9 publications

References 40 publications

Effects of Human Endothelial Progenitor Cell and Its Conditioned Medium on Oocyte Development and Subsequent Embryo Development

Effects of Human Endothelial Progenitor Cell and Its Conditioned Medium on Oocyte Development and Subsequent Embryo Development

Effects of dexamethasone on growth, viability and ultrastructure of bovine secondary follicles cultured in vitro

Why Is It So Difficult To Have Competent Oocytes from In vitro Cultured Preantral Follicles?

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