Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources during<i>in vitro</i>culture

Gomes, Debir Soares; Aragão, L B; Neto, M F Lima; Barroso, P.A.A.; Paulino, Laís R F M; Silva, B R; Souza, Ana Liza Paz; Vasconcelos, G. L. de; Silva, A W B; Silva, J R V

doi:10.1017/s0967199419000583

Cited by 6 publications

(5 citation statements)

References 30 publications

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“…Notably, its use has been consolidated over recent years due to its longer circulatory half-life, enhanced trans-species effect, lower cost, and greater commercial availability compared with FSH [54,55]. Despite these numerous advantages, no evidence has been collected to date documenting eCG's influence on the promotion of in vitro folliculogenesis.…”

Section: Discussionmentioning

confidence: 99%

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Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development

Berardino

Peserico

Capacchietti

et al. 2021

IJMS

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The use of assisted reproductive technologies (ART) still requires strategies through which to maximize individual fertility chances. In vitro folliculogenesis (ivF) may represent a valid option to convey the large source of immature oocytes in ART. Several efforts have been made to set up ivF cultural protocols in medium-sized mammals, starting with the identification of the most suitable gonadotropic stimulus. In this study, Equine Chorionic Gonadotropin (eCG) is proposed as an alternative to Follicle Stimulating Hormone (FSH) based on its long superovulation use, trans-species validation, long half-life, and low costs. The use of 3D ivF on single-ovine preantral (PA) follicles allowed us to compare the hormonal effects and to validate their influence under two different cultural conditions. The use of eCG helped to stimulate the in vitro growth of ovine PA follicles by maximizing its influence under FBS-free medium. Higher performance of follicular growth, antrum formation, steroidogenic activity and gap junction marker expression were recorded. In addition, eCG, promoted a positive effect on the germinal compartment, leading to a higher incidence of meiotic competent oocytes. These findings should help to widen the use of eCG to ivF as a valid and largely available hormonal support enabling a synchronized in vitro follicle and oocyte development.

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Section: Discussionmentioning

confidence: 99%

“…Equine Chorionic Gonadotropin is a placental glycoprotein obtained from the serum of pregnant mares. It is widely used as a valid substitute for species-specific FSH in female mammals due to its longer circulatory half-life, its FSH like-activity, its easier use as a trans-species hormone, and its limited costs [54,55].…”

Section: Introductionmentioning

confidence: 99%

Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development

Berardino

Peserico

Capacchietti

et al. 2021

IJMS

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“…After 18 days of culture, the follicles were fixed (n = 8, per treatment) for 24 h at room temperature in 4% paraformaldehyde in phosphate-buffered saline (PBS; pH 7.4). After fixation, follicles were dehydrated in a graded series of ethanol concentrations (70, 80, 90 and 100%), clarified with xylol, and embedded in paraffin (Gomes et al, 2020). For each group of follicles, 5-μm sections were mounted on slides and stained with eosin and haematoxylin.…”

Section: Histological Analysis Of Cultured Secondary Folliclesmentioning

confidence: 99%

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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“…Nowadays, the desire to minimize the introduction of xenogeneic proteins or pathogens and to standardize cellular products intended for clinical application has driven the search for alternatives to FBS. Bovine serum albumin (BSA), as the most abundant protein in plasma, is probably better than FBS owing to lower xenogeneic proteins or pathogens [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Optimized study of anin vitro3D culture of preantral follicles in mice

Ren

Zhang

et al. 2023

J Vet Sci

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Background In vitro culture of preantral follicles is a promising technology for fertility preservation. Objectives This study aims to investigate an optimized three-dimensional (3D) fetal bovine serum (FBS)-free preantral follicle culture system having a simple and easy operation. Methods The isolated follicles from mouse ovaries were randomly divided in an ultra-low attachment 96-well plates supplement with FBS or bovine serum albumin (BSA) culture or encapsulated with an alginate supplement with FBS or BSA culture. Meanwhile, estradiol (E 2 ) concentration was assessed through enzyme-linked immunosorbent assay of culture supernatants. The diameter of follicular growth was measured, and the lumen of the follicle was photographed. Spindle microtubules of oocytes were detected via immunofluorescence. The ability of oocytes to fertilize was assessed using in vitro fertilization. Results The diameters were larger for the growing secondary follicles cultured in ultra-low attachment 96-well plates than in the alginate gel on days 6, 8, and 10 ( p < 0.05). Meanwhile, the E 2 concentration in the BSA-supplemented medium was significantly higher in the alginate gel than in the other three groups on days 6 and 8 ( p < 0.05), and the oocytes in the FBS-free system could complete meiosis and fertilization in vitro . Conclusions The present study furnishes insights into the mature oocytes obtained from the 3D culture of the preantral follicle by using ultra-low attachment 96-well plate with an FBS-free system in vitro and supports the clinical practices to achieve competent, mature oocytes for in vitro fertilization.

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Supplementation of culture medium with knockout serum replacement improves the survival of bovine secondary follicles when compared with other protein sources duringin vitroculture

Cited by 6 publications

References 30 publications

Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development

Equine Chorionic Gonadotropin as an Effective FSH Replacement for In Vitro Ovine Follicle and Oocyte Development

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

Optimized study of anin vitro3D culture of preantral follicles in mice

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