Abstract
To compare the effects of two-(2D, microplate) and three-dimensional (3D, alginate) culture systems on the in vitro growth of small antral follicles in cattle, individual follicles were separately cultured in the two culture systems for 8 days. Half of the culture medium was replaced by fresh medium every 2 days; the former medium was used to assess the amount of follicular hormone secretion using ELISA. Individual follicle morphology, diameter, and survival rate were recorded every alternate day. The results showed that in 4 days, there was no significant difference between the two systems, except that the growth rate of follicles in 2D system was relatively faster. After 4 days, estradiol concentration in 3D system was higher than that in 2D system. However, progesterone concentration was lower than that in the 2D system. The survival rate and oocyte quality of follicles in 2D system were significantly lower than those in 3D system on day 8. The follicle diameter slightly increased (30-60 μm) in the entire process. Taken together, for in vitro culture of follicles within 4 days, the 2D culture system is more suitable. However, when the culture duration is >4 days, the 3D culture system is more suitable.
Highlights
Understanding the regulatory mechanism of ovarian follicle growth and atresia is clinically significant in the selection of high-quality germ cells for assisted reproduction
Culture systems have been established for culturing preantral follicles, such as those found in sheep (Praveen Chakravarthi et al 2015), goat (Silva et al 2015), buffalo (Gupta et al 2008), pig (Wu et al 2001), and cattle (Petro et al 2010)
225 small antral follicles were divided into three categories: group I, in which 71 follicles have an oocyte surrounded by cumulus cells, with intact mural granulosa cells and limited by an intact basement membrane and outer thecal-stromal layer rich in blood vessels (Fig. 1 aI); group II, in which 76 follicles have a disrupted architecture in the mural granulosa cell layer inside the basal membrane (Fig. 1 aII); and group III, in which 78 follicles, the mural granulosa cell layers inside
Summary
Understanding the regulatory mechanism of ovarian follicle growth and atresia is clinically significant in the selection of high-quality germ cells for assisted reproduction. Many follicles are activated into the growth phase from fetal life through reproductive age. In vitro culture is a great way to increase the utilization rate of follicles (Gomes et al 2015, Haag et al 2013). Various murine in vitro culture systems have been successfully established for the growth of mature mouse oocytes from primordial follicles, in vitro fertilization, and development of fertile progeny after embryo transfer (Itami et al 2014). Culture systems have been established for culturing preantral follicles, such as those found in sheep (Praveen Chakravarthi et al 2015), goat (Silva et al 2015), buffalo (Gupta et al 2008), pig (Wu et al 2001), and cattle (Petro et al 2010).
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