![]() Additionally, these methods have made it easier to study the mechanics, extracellular matrix, and gene expression during folliculogenesis. ![]() Preantral mouse, nonhuman primate, and human follicles have been grown in in vitro systems based on alginate, with mice follicles found to produce oocytes capable of fertilization and live birth. Alginate is a polysaccharide polymer generated from algae that has been used in several tissue growth applications. Furthermore, alginate hydrogels are a common tissue engineering scaffold that replicate the ovaries by providing an ideal 3D environment and encouraging somatic cell–egg interactions to improve oocyte development. Maintaining the spherical form of ovarian follicles and improving cell viability and growth in in vitro culture are the two major benefits of 3D follicle cultures. The 3D system has also been used successfully in a wider range of species. The 3D system has attracted more attention in recent years for follicle culture than has the 2D system. ĭue to the lack of correlation between the cultivated follicle microenvironment and in vivo environments, unlike the 3D system, the 2D system does not sustain follicle integrity. These methods enable the development of immature follicles in vitro, resulting in the production of oocytes that can develop, be fertilized, and give rise to live birth in mice. In this regard, 2-dimensional (2D) and 3-dimensional (3D) culture systems have been developed. ![]() In vitro production of isolated immature ovarian follicles is a promising option for fertility preservation in adult or prepubertal patients with cancers that may migrate to the ovaries. Isolated and in vitro cultivation of ovarian follicles have been investigated as an alternative approach to overcome the limitations of ovarian tissue transplantation. In vitro follicle cultures can be used with cryopreserved ovarian tissue to produce mature eggs and, thereafter, embryos. Patients with cancer can choose to freeze their ovarian tissue prior to treatment however, autotransplant of cryopreserved ovarian tissue following cancer treatment poses the risk of reintroducing cancer cells. These treatments reduce the follicle pool and cause early ovarian failure. Cancer treatments, including chemotherapy and radiotherapy, can cause infertility in girls and women. In vitro ovarian follicle cultivation systems have enabled the development of fertility preservation methods for patients with cancer. A 3D culture system using an alginate matrix and supplemented with BMP-15 effectively improves the outcomes of in vitro ovarian follicle culture. Intracellular reactive oxygen species levels were higher and glutathione levels were lower in mature oocytes from the in vitro culture than in mature oocytes from an in vivo control. The result for E2 tended toward significance on day 12. The secretion of P4 was significantly higher on days 6, 8, and 10 in follicles cultured in BMP-15-supplemented medium. The percentage of ovulated metaphase II oocytes retrieved from follicles cultured in BMP-15-supplemented medium was greater than that of oocytes retrieved from follicles cultured in regular medium. On day 12, the follicle diameter, follicle survival rate, and antrum formation rate were significantly higher for follicles cultured in BMP-15-supplemented medium than those cultured in regular medium. ![]() The in vitro development of ovarian follicles was evaluated. Multilayered secondary follicles were encapsulated in a 0.5% alginate matrix and cultured in a 3D culture system supplemented with bone morphogenetic protein 15 (BMP-15 15 ng/mL) for 12 days. However, mature oocytes retrieved in vitro were subject to hypoxia conditions. Regarding observations of follicle growth, in vitro culture using 0.5% alginate supplemented with 15 ng/mL BMP-15 can promote the production of mature oocytes and increase steroidogenesis. This protein functions primarily by attaching to its receptor on the surface of granulosa cells, which are also essential for the development of ovarian follicles. BMP-15 is a member of the superfamily of transforming growth factor-beta (TGF-β), a protein that is necessary for ovarian follicular growth. Typically employed for follicle culture, alginate prepared at a concentration of 0.5% w/ v led to considerable developmental competence and steroidogenesis. Alginate is used in in vitro culture systems to mimic the natural ovarian environment by maintaining the 3D follicular architecture, cell–cell interactions, and paracrine signals underlying direct follicle development. Multilayered secondary follicles were encapsulated and cultured for 12 days in a 3D culture system by using alginate supplemented with bone morphogenetic protein 15 (BMP-15).
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