In vitro growth and development of isolated secondary follicles from vitrified caprine ovarian cortex

Abstract

The aim of this study was to evaluate the viability, antrum formation and in vitro development of isolated secondary follicles from vitrified caprine ovarian cortex in a medium previously established for fresh isolated secondary follicles, in the absence (α-minimum essential medium (α-MEM+) alone) or presence of FSH and vascular endothelial growth factor (VEGF; α-MEM++FSH+VEGF). Ovarian fragments were distributed among five treatments (T1 to T5): fresh follicles were fixed immediately (T1), follicles from fresh tissue were cultured in vitro in α-MEM+ (T2) or α-MEM++FSH+VEGF (T3) and follicles from vitrified tissue were cultured in vitro in α-MEM+ (T4) or α-MEM++FSH+VEGF (T5). After 6 days of culture, treated follicles (T2, T3, T4 and T5) were evaluated for morphology, viability and follicular development (growth, antrum formation and proliferation of granulosa cells by Ki67 and argyrophilic nucleolar organiser region (AgNOR) staining). The levels of reactive oxygen species (ROS) in the culture media were also assessed. Overall, morphology of vitrified follicles was altered (P<0.05) compared with the fresh follicles. Follicular viability, antrum formation and ROS were similar between treatments (P>0.05). The average overall and daily follicular growth was highest (P<0.05) in T3. Granulosa cells in all treatments (T1, T2, T3, T4 and T5) stained positive for Ki67. However, fresh follicles from T3 had significantly higher AgNOR staining (P<0.05) compared with follicles of T1, T2, T4 and T5. In conclusion, secondary follicles can be isolated from vitrified and warmed ovarian cortex and survive and form an antrum when growing in an in vitro culture for 6 days.

Oncofertility Approach in Females With Leukemia From the Reproductive Perspective

“…54 Moreover, techniques like in vitro growth of harvested follicles, oocyte generation using ovarian tissue xenografts in animal models, as well as stem cell (e.g., mesenchymal) utilization, might be available in the future with limited ethical dilemmas. [61][62][63][64] Additionally, understanding of the gonadotoxic mechanisms of commonly used chemotherapeutic agents has enabled the development of preventive adjuvant pharmacotherapy. 46 There is now a range of agents attempting to protect the ovary against chemotherapy by preventing direct loss and accelerated activation of primordial follicles, as well as follicular atresia.…”

In vitro growth of the ovarian follicle: taking stock of advances in research

“…Follicles produced in vitro are small compared to follicles produced in vivo (Xiao et al, 2015;Rodrigues et al, 2015). Rodents and mammals are the most prevalent models, approximately one-fifth of the studies use human follicles (Xiao et al, 2015;Telfer et al, 2008), and other mammalian follicles, like the Rhesus monkey (Rodrigues et al, 2015;Xu et al, 2009a;Peluffo et al, 2010;Xu et al, 2011a;Hornick et al, 2012;Xu et al, 2013;Xu et al, 2015b;Xu et al, 2018;Baba et al, 2017); baboon (Xu et al, 2011b); bovine (Yamamoto et al, 1999;Rossetto et al, 2013a;Araújo et al, 2015); ovine (Arunakumari et al, 2010;Muruvi et al, 2005); caprine (Rossetto et al, 2013a;Ferreira et al, 2018;Silva et al, 2015;Magalhães et al, 2011); swine (Hirao et al, 1994;Wu et al, 2001); cats (Songsasen et al, 2017;Thongkittidilok et al, 2018); dogs (Songsasen et al, 2011); horses (Haag et al, 2013); wildcats (Wiedemann et al, 2013).…”

“…In most rodent follicle culture studies, prepubertal follicles have been used, and in less than 30% of adult follicles (Diaz et al, 2007;Simon et al, 2020). Young animals of reproductive age have been used in studies of mammalian follicles such as sheep (Thomas et al, 2003;Arunakumari et al, 2010), goats (Ferreira et al, 2018;Magalhães et al, 2011), and cattle (Gutierrez et al, 2000;Itoh et al, 2002;Araújo et al, 2014a;2015). Prepubertal follicles and smaller follicles have been used to evaluate the use of FSH supplementation in cattle and sheep (Wandji et al, 1996;Cecconi et al, 1999;Muruvi et al, 2005).…”

Effect of animal-sourced bioactive peptides on the in vitro development of mouse preantral follicles

“…However, the production and accumulation of reactive oxygen species (ROS) from cellular metabolic processes in the developing follicles affects the development and survival of follicles during long-term culture [7]. Indeed, when excessive ROS levels overwhelm the balance of the cellular antioxidant defense system, oxidative stress (OS) occurs [8,9], resulting in direct or indirect ROSmediated damage to nucleic DNA, proteins and lipids, thereby altering their functions and resulting in the abnormal gene expression patterns, DNA double strand break (DSB) repair, chromosomal errors and follicular development [10][11][12][13].…”